C e ll, Vo l.

1 0 0 , 5 7 7 0 , J a n u a ry 7 , 2 0 0 0 , C o p yrig h t 2 0 0 0 b y C e ll P re ss
The Hallmarks of Cancer Review
e vo lve p ro g re ssive ly fro m n o rm a lc y via a se rie s o f p re - Douglas Hanahan* and Robert A. Weinberg

* D e p a rtm e n t o f B io c h e m istry a n d B io p h ysic s a n d m a lig n a n t sta te s in to in va sive c a n c e rs F o u ld s, 1 9 5 4 .

T h e se o b se rva tio n s h a ve b e e n re n d e re d m o re c o n - H o rm o n e R e se a rc h I n stitu te
U n ive rsity o f C a lifo rn ia a t S a n F ra n c isc o c re te b y a la rg e b o d y o f w o rk in d ic a tin g th a t th e g e -
n o m e s o f tu m o r c e lls a re in va ria b ly a lte re d a t m u ltip le S a n F ra n c isc o , C a lifo rn ia 9 4 1 4 3

Wh ite h e a d I n stitu te fo r B io m e d ic a l R e se a rc h a n d site s, h a vin g su ffe re d d isru p tio n th ro u g h le sio n s a s su b -

tle a s p o in t m u ta tio n s a n d a s o b vio u s a s c h a n g e s in D e p a rtm e n t o f B io lo g y
M a ssa c h u se tts I n stitu te o f T e c h n o lo g y c h ro m o so m e c o m p le m e n t e . g . , K in zle r a n d Vo g e lste in ,
1 9 9 6 . T ra n sfo rm a tio n o f c u ltu re d c e lls is itse lf a C a m b rid g e , M a ssa c h u se tts 0 2 1 4 2
m u ltiste p p ro c e ss: ro d e n t c e lls re q u ire a t le a st tw o in tro -
d u c e d g e n e tic c h a n g e s b e fo re th e y a c q u ire tu m o rig e n ic
c o m p e te n c e , w h ile th e ir h u m a n c o u n te rp a rts a re m o re A fte r a q u a rte r c e n tu ry o f ra p id a d va n c e s, c a n c e r re -
d iffic u lt to tra n sfo rm H a h n e t a l. , 1 9 9 9 . T ra n sg e n ic se a rc h h a s g e n e ra te d a ric h a n d c o m p le x b o d y o f k n o w l-
m o d e ls o f tu m o rig e n e sis h a ve re p e a te d ly su p p o rte d th e e d g e , re ve a lin g c a n c e r to b e a d ise a se in vo lvin g d y-
c o n c lu sio n th a t tu m o rig e n e sis in m ic e in vo lve s m u ltip le n a m ic c h a n g e s in th e g e n o m e . T h e fo u n d a tio n h a s b e e n
ra te -lim itin g ste p s B e rg e rs e t a l. , 1 9 9 8 ; se e Oncogene, se t in th e d isc o ve ry o f m u ta tio n s th a t p ro d u c e o n c o -
1 9 9 9 , R . D e P in h o a n d T . E . J a c k s, vo lu m e 1 8 [3 8 ], p p . g e n e s w ith d o m in a n t g a in o f fu n c tio n a n d tu m o r su p -
5 2 4 8 5 3 6 2 . T a k e n to g e th e r, o b se rva tio n s o f h u m a n p re sso r g e n e s w ith re c e ssive lo ss o f fu n c tio n ; b o th
c a n c e rs a n d a n im a l m o d e ls a rg u e th a t tu m o r d e ve lo p - c la sse s o f c a n c e r g e n e s h a ve b e e n id e n tifie d th ro u g h
m e n t p ro c e e d s via a p ro c e ss fo rm a lly a n a lo g o u s to D a r- th e ir a lte ra tio n in h u m a n a n d a n im a l c a n c e r c e lls a n d
w in ia n e vo lu tio n , in w h ic h a su c c e ssio n o f g e n e tic b y th e ir e lic ita tio n o f c a n c e r p h e n o typ e s in e x p e rim e n ta l
c h a n g e s, e a c h c o n fe rrin g o n e o r a n o th e r typ e o f g ro w th m o d e ls B ish o p a n d We in b e rg , 1 9 9 6 .
a d va n ta g e , le a d s to th e p ro g re ssive c o n ve rsio n o f n o r- S o m e w o u ld a rg u e th a t th e se a rc h fo r th e o rig in a n d
m a l h u m a n c e lls in to c a n c e r c e lls F o u ld s, 1 9 5 4 ; N o w e ll, tre a tm e n t o f th is d ise a se w ill c o n tin u e o ve r th e n e x t
1 9 7 6 . q u a rte r c e n tu ry in m u c h th e sa m e m a n n e r a s it h a s in
th e re c e n t p a st, b y a d d in g fu rth e r la ye rs o f c o m p le x ity
to a sc ie n tific lite ra tu re th a t is a lre a d y c o m p le x a lm o st AnEnumerationof the Traits
b e yo n d m e a su re . B u t w e a n tic ip a te o th e rw ise : th o se T h e b a rrie rs to d e ve lo p m e n t o f c a n c e r a re e m b o d ie d
re se a rc h in g th e c a n c e r p ro b le m w ill b e p ra c tic in g a d ra - in a te le o lo g y: c a n c e r c e lls h a ve d e fe c ts in re g u la to ry
m a tic a lly d iffe re n t typ e o f sc ie n c e th a n w e h a ve e x p e ri- c irc u its th a t g o ve rn n o rm a l c e ll p ro life ra tio n a n d h o m e o -
e n c e d o ve rth e p a st 2 5 ye a rs. S u re ly m u c h o f th is c h a n g e sta sis. T h e re a re m o re th a n 1 0 0 d istin c t typ e s o f c a n c e r,
w ill b e a p p a re n t a t th e te c h n ic a l le ve l. B u t u ltim a te ly, a n d su b typ e s o f tu m o rs c a n b e fo u n d w ith in sp e c ific
th e m o re fu n d a m e n ta l c h a n g e w ill b e c o n c e p tu a l. o rg a n s. T h is c o m p le x ity p ro vo k e s a n u m b e r o f q u e s-
We fo re se e c a n c e r re se a rc h d e ve lo p in g in to a lo g ic a l tio n s. H o w m a n y d istin c t re g u la to ry c irc u its w ith in e a c h
sc ie n c e , w h e re th e c o m p le x itie s o f th e d ise a se , d e - typ e o f ta rg e t c e ll m u st b e d isru p te d in o rd e r fo r su c h
sc rib e d in th e la b o ra to ry a n d c lin ic , w ill b e c o m e u n d e r- a c e ll to b e c o m e c a n c e ro u s? D o e s th e sa m e se t o f
sta n d a b le in te rm s o f a sm a ll n u m b e r o f u n d e rlyin g p rin - c e llu la r re g u la to ry c irc u its su ffe r d isru p tio n in th e c e lls
c ip le s. S o m e o f th e se p rin c ip le s a re e ve n n o w in th e o f th e d isp a ra te n e o p la sm s a risin g in th e h u m a n b o d y?
m id st o f b e in g c o d ifie d . We d isc u ss o n e se t o f th e m in Wh ic h o f th e se c irc u its o p e ra te o n a c e ll-a u to n o m o u s
th e p re se n t e ssa y: ru le s th a t g o ve rn th e tra n sfo rm a tio n b a sis, a n d w h ic h a re c o u p le d to th e sig n a ls th a t c e lls
o f n o rm a l h u m a n c e lls in to m a lig n a n t c a n c e rs. We su g - re c e ive fro m th e ir su rro u n d in g m ic ro e n viro n m e n t w ith in
g e st th a t re se a rc h o ve r th e p a st d e c a d e s h a s re ve a le d a tissu e ? C a n th e la rg e a n d d ive rse c o lle c tio n o f c a n c e r-
a sm a ll n u m b e r o f m o le c u la r, b io c h e m ic a l, a n d c e llu la r a sso c ia te d g e n e s b e tie d to th e o p e ra tio n s o f a sm a ll
tra its a c q u ire d c a p a b ilitie s sh a re d b y m o st a n d p e r- g ro u p o f re g u la to ry c irc u its?
h a p s a ll typ e s o f h u m a n c a n c e r. O u rfa ith in su c h sim p lifi- We su g g e st th a t th e va st c a ta lo g o f c a n c e r c e ll g e n o -
c a tio n d e rive s d ire c tly fro m th e te a c h in g s o f c e ll b io lo g y typ e s is a m a n ife sta tio n o f six e sse n tia l a lte ra tio n s in c e ll
th a t virtu a lly a ll m a m m a lia n c e lls c a rry a sim ila r m o le c u - p h ysio lo g y th a t c o lle c tive ly d ic ta te m a lig n a n t g ro w th
la r m a c h in e ry re g u la tin g th e ir p ro life ra tio n , d iffe re n tia - F ig u re 1 : se lf-su ffic ie n c y in g ro w th sig n a ls, in se n sitivity
tio n , a n d d e a th . to g ro w th -in h ib ito ry a n tig ro w th sig n a ls, e va sio n o f p ro -
S e ve ra l lin e s o f e vid e n c e in d ic a te th a t tu m o rig e n e sis g ra m m e d c e ll d e a th a p o p to sis , lim itle ss re p lic a tive
in h u m a n s is a m u ltiste p p ro c e ss a n d th a t th e se ste p s p o te n tia l, su sta in e d a n g io g e n e sis, a n d tissu e in va sio n
re fle c t g e n e tic a lte ra tio n s th a t d rive th e p ro g re ssive a n d m e ta sta sis. E a c h o f th e se p h ysio lo g ic c h a n g e s
tra n sfo rm a tio n o f n o rm a l h u m a n c e lls in to h ig h ly m a lig - n o ve l c a p a b ilitie s a c q u ire d d u rin g tu m o r d e ve lo p m e n t
n a n t d e riva tive s. M a n y typ e s o f c a n c e rs a re d ia g n o se d re p re se n ts th e su c c e ssfu l b re a c h in g o f a n a n tic a n c e r
in th e h u m a n p o p u la tio n w ith a n a g e -d e p e n d e n t in c i- d e fe n se m e c h a n ism h a rd w ire d in to c e lls a n d tissu e s.
d e n c e im p lic a tin g fo u r to se ve n ra te -lim itin g , sto c h a stic We p ro p o se th a t th e se six c a p a b ilitie s a re sh a re d in
e ve n ts R e n a n , 1 9 9 3 . P a th o lo g ic a l a n a lyse s o f a n u m b e r c o m m o n b y m o st a n d p e rh a p s a ll typ e s o f h u m a n tu -
o f o rg a n site s re ve a l le sio n s th a t a p p e a r to re p re se n t m o rs. T h is m u ltip lic ity o f d e fe n se s m a y e x p la in w h y c a n -
c e r is re la tive ly ra re d u rin g a n a ve ra g e h u m a n life tim e . th e in te rm e d ia te ste p s in a p ro c e ss th ro u g h w h ic h c e lls
C e ll
5 8
A c q u ire d G S a u to n o m y w a s th e first o f th e six c a p a b ili-
tie s to b e c le a rly d e fin e d b y c a n c e r re se a rc h e rs, in la rg e
p a rt b e c a u se o f th e p re va le n c e o f d o m in a n t o n c o g e n e s
th a t h a ve b e e n fo u n d to m o d u la te it. T h re e c o m m o n
m o le c u la r stra te g ie s fo r a c h ie vin g a u to n o m y a re e vi-
d e n t, in vo lvin g a lte ra tio n o f e x tra c e llu la r g ro w th sig n a ls,
o f tra n sc e llu la r tra n sd u c e rs o f th o se sig n a ls, o r o f in tra -
c e llu la r c irc u its th a t tra n sla te th o se sig n a ls in to a c tio n .
Wh ile m o st so lu b le m ito g e n ic g ro w th fa c to rs G F s a re
m a d e b y o n e c e ll typ e in o rd e r to stim u la te p ro life ra tio n
o f a n o th e r th e p ro c e ss o f h e te ro typ ic sig n a lin g m a n y
c a n c e r c e lls a c q u ire th e a b ility to syn th e size G F s to
w h ic h th e y a re re sp o n sive , c re a tin g a p o sitive fe e d b a c k
sig n a lin g lo o p o fte n te rm e d a u to c rin e stim u la tio n F e d i
e t a l. , 1 9 9 7 . C le a rly, th e m a n u fa c tu re o f a G F b y a c a n c e r
c e ll o b via te s d e p e n d e n c e o n G F s fro m o th e r c e lls w ith in
th e tissu e . T h e p ro d u c tio n o f P D G F p la te le t-d e rive d
g ro w th fa c to r a n d T G F tu m o r g ro w th fa c to r b y
g lio b la sto m a s a n d sa rc o m a s, re sp e c tive ly, a re tw o illu s-
tra tive e x a m p le s F e d i e t a l. , 1 9 9 7 .
T h e c e ll su rfa c e re c e p to rs th a t tra n sd u c e g ro w th -
stim u la to ry sig n a ls in to th e c e ll in te rio r a re th e m se lve s
ta rg e ts o f d e re g u la tio n d u rin g tu m o r p a th o g e n e sis. G F
re c e p to rs, o fte n c a rryin g tyro sin e k in a se a c tivitie s in
th e ir c yto p la sm ic d o m a in s, a re o ve re x p re sse d in m a n y
c a n c e rs. R e c e p to r o ve re x p re ssio n m a y e n a b le th e c a n -
c e r c e ll to b e c o m e h yp e rre sp o n sive to a m b ie n t le ve ls
F ig u re 1 . A c q u ire d C a p a b ilitie s o f C a n c e r
o f G F th a t n o rm a lly w o u ld n o t trig g e r p ro life ra tio n F e d i
We su g g e st th a t m o st if n o t a ll c a n c e rs h a ve a c q u ire d th e sa m e se t
e t a l. , 1 9 9 7 . F o r e x a m p le , th e e p id e rm a l G F re c e p to r o f fu n c tio n a l c a p a b ilitie s d u rin g th e ir d e ve lo p m e n t, a lb e it th ro u g h
va rio u s m e c h a n istic stra te g ie s. E G F -R /erbB is u p re g u la te d in sto m a c h , b ra in , a n d
b re a st tu m o rs, w h ile th e H E R 2 /neu re c e p to r is o ve re x -
p re sse d in sto m a c h a n d m a m m a ry c a rc in o m a s S la m o n
e t a l. , 1 9 8 7 ; Ya rd e n a n d U llric h , 1 9 8 8 . A d d itio n a lly, g ro ss
We d e sc rib e e a c h c a p a b ility in tu rn b e lo w , illu stra te w ith
o ve re x p re ssio n o f G F re c e p to rs c a n e lic it lig a n d -in d e -
a fe w e x a m p le s its fu n c tio n a l im p o rta n c e , a n d in d ic a te
p e n d e n t sig n a lin g D iF io re e t a l. , 1 9 8 7 . L ig a n d -in d e p e n -
stra te g ie s b y w h ic h it is a c q u ire d in h u m a n c a n c e rs.
d e n t sig n a lin g c a n a lso b e a c h ie ve d th ro u g h stru c tu ra l
a lte ra tio n o f re c e p to rs; fo r e x a m p le , tru n c a te d ve rsio n s
Acquired Capability: Self-Sufficiency
o f th e E G F re c e p to r la c k in g m u c h o f its c yto p la sm ic
inGrowthSignals
d o m a in fire c o n stitu tive ly F e d i e t a l. , 1 9 9 7 .
N o rm a l c e lls re q u ire m ito g e n ic g ro w th sig n a ls G S b e -
C a n c e r c e lls c a n a lso sw itc h th e typ e s o f e x tra c e llu la r
fo re th e y c a n m o ve fro m a q u ie sc e n t sta te in to a n a c tive
m a trix re c e p to rs in te g rin s th e y e x p re ss, fa vo rin g o n e s
p ro life ra tive sta te . T h e se sig n a ls a re tra n sm itte d in to th e
th a t tra n sm it p ro g ro w th sig n a ls L u k a sh e v a n d We rb ,
c e ll b y tra n sm e m b ra n e re c e p to rs th a t b in d d istin c tive
1 9 9 8 ; G ia n c o tti a n d R u o sla h ti, 1 9 9 9 . T h e se b ifu n c tio n a l,
c la sse s o f sig n a lin g m o le c u le s: d iffu sib le g ro w th fa c -
h e te ro d im e ric c e ll su rfa c e re c e p to rs p h ysic a lly lin k c e lls
to rs, e x tra c e llu la r m a trix c o m p o n e n ts, a n d c e ll-to -c e ll
to e x tra c e llu la r su p e rstru c tu re s k n o w n a s th e e x tra c e llu -
a d h e sio n /in te ra c tio n m o le c u le s. T o o u r k n o w le d g e , n o
la r m a trix E C M . S u c c e ssfu l b in d in g to sp e c ific m o ie tie s
typ e o f n o rm a l c e ll c a n p ro life ra te in th e a b se n c e o f
o f th e E C M e n a b le s th e in te g rin re c e p to rs to tra n sd u c e
su c h stim u la to ry sig n a ls. M a n y o f th e o n c o g e n e s in th e
sig n a ls in to th e c yto p la sm th a t in flu e n c e c e ll b e h a vio r,
c a n c e r c a ta lo g a c t b y m im ic k in g n o rm a l g ro w th sig n a l-
ra n g in g fro m q u ie sc e n c e in n o rm a l tissu e to m o tility,
in g in o n e w a y o r a n o th e r.
re sista n c e to a p o p to sis, a n d e n tra n c e in to th e a c tive
D e p e n d e n c e o n g ro w th sig n a lin g is a p p a re n t w h e n
c e ll c yc le . C o n ve rse ly, th e fa ilu re o f in te g rin s to fo rg e
p ro p a g a tin g n o rm a l c e lls in c u ltu re , w h ic h typ ic a lly p ro -
th e se e x tra c e llu la r lin k s c a n im p a ir c e ll m o tility, in d u c e
life ra te o n ly w h e n su p p lie d w ith a p p ro p ria te d iffu sib le
a p o p to sis, o r c a u se c e ll c yc le a rre st G ia n c o tti a n d R u -
m ito g e n ic fa c to rs a n d a p ro p e r su b stra tu m fo r th e ir in te -
o sla h ti, 1 9 9 9 . B o th lig a n d -a c tiva te d G F re c e p to rs a n d
g rin s. S u c h b e h a vio r c o n tra sts stro n g ly w ith th a t o f tu -
p ro g ro w th in te g rin s e n g a g e d to e x tra c e llu la r m a trix
m o r c e lls, w h ic h in va ria b ly sh o w a g re a tly re d u c e d
c o m p o n e n ts c a n a c tiva te th e S O S -R a s-R a f-M A P k in a se
d e p e n d e n c e o n e x o g e n o u s g ro w th stim u la tio n . T h e c o n -
p a th w a y A p lin e t a l. , 1 9 9 8 ; G ia n c o tti a n d R u o sla h ti,
c lu sio n is th a t tu m o r c e lls g e n e ra te m a n y o f th e ir o w n
1 9 9 9 .
g ro w th sig n a ls, th e re b y re d u c in g th e ir d e p e n d e n c e o n T h e m o st c o m p le x m e c h a n ism s o f a c q u ire d G S a u to n -
stim u la tio n fro m th e ir n o rm a l tissu e m ic ro e n viro n m e n t. o m y d e rive fro m a lte ra tio n s in c o m p o n e n ts o f th e d o w n -
T h is lib e ra tio n fro m d e p e n d e n c e o n e x o g e n o u sly d e - stre a m c yto p la sm ic c irc u itry th a t re c e ive s a n d p ro -
rive d sig n a ls d isru p ts a c ritic a lly im p o rta n t h o m e o sta tic c e sse s th e sig n a ls e m itte d b y lig a n d -a c tiva te d G F
m e c h a n ism th a t n o rm a lly o p e ra te s to e n su re a p ro p e r re c e p to rs a n d in te g rin s. T h e S O S -R a s-R a f-M A P K c a s-
c a d e p la ys a c e n tra l ro le h e re . I n a b o u t 2 5 % o f h u m a n b e h a vio r o f th e va rio u s c e ll typ e s w ith in a tissu e .
R e vie w
5 9
F ig u re 2 . T h e E m e rg e n t I n te g ra te d C irc u it o f th e C e ll
P ro g re ss in d isse c tin g sig n a lin g p a th w a ys h a s b e g u n to la y o u t a c irc u itry th a t w ill lik e ly m im ic e le c tro n ic in te g ra te d c irc u its in c o m p le x ity
a n d fin e sse , w h e re tra n sisto rs a re re p la c e d b y p ro te in s e . g . , k in a se s a n d p h o sp h a ta se s a n d th e e le c tro n s b y p h o sp h a te s a n d lip id s, a m o n g
o th e rs. I n a d d itio n to th e p ro to typ ic a l g ro w th sig n a lin g c irc u it c e n te re d a ro u n d R a s a n d c o u p le d to a sp e c tru m o f e x tra c e llu la r c u e s, o th e r
c o m p o n e n t c irc u its tra n sm it a n tig ro w th a n d d iffe re n tia tio n sig n a ls o r m e d ia te c o m m a n d s to live o r d ie b y a p o p to sis. A s fo r th e g e n e tic
re p ro g ra m m in g o f th is in te g ra te d c irc u it in c a n c e r c e lls, so m e o f th e g e n e s k n o w n to b e fu n c tio n a lly a lte re d a re h ig h lig h te d in re d .
tu m o rs, R a s p ro te in s a re p re se n t in stru c tu ra lly a lte re d m u ltip le c e ll b io lo g ic a l e ffe c ts. F o r e x a m p le , th e d ire c t
in te ra c tio n o f th e R a s p ro te in w ith th e su rviva l-p ro m o t- fo rm s th a t e n a b le th e m to re le a se a flu x o f m ito g e n ic
sig n a ls in to c e lls, w ith o u t o n g o in g stim u la tio n b y th e ir in g P I 3 k in a se e n a b le s g ro w th sig n a ls to c o n c u rre n tly
e vo k e su rviva l sig n a ls w ith in th e c e ll D o w n w a rd , 1 9 9 8 . n o rm a l u p stre a m re g u la to rs M e d e m a a n d B o s, 1 9 9 3 .
We su sp e c t th a t g ro w th sig n a lin g p a th w a ys su ffe r Wh ile a c q u isitio n o f g ro w th sig n a lin g a u to n o m y b y
c a n c e r c e lls is c o n c e p tu a lly sa tisfyin g , it is a lso to o d e re g u la tio n in a ll h u m a n tu m o rs. A lth o u g h th is p o in t
is h a rd to p ro ve rig o ro u sly a t p re se n t, th e c lu e s a re sim p listic . We h a ve tra d itio n a lly e x p lo re d tu m o r g ro w th
b y fo c u sin g o u r e x p e rim e n ta l a tte n tio n s o n th e g e n e ti- a b u n d a n t H u n te r, 1 9 9 7 . F o r e x a m p le , in th e b e st stu d -
ie d o f tu m o rs h u m a n c o lo n c a rc in o m a s a b o u t h a lf c a lly d e ra n g e d c a n c e r c e lls F ig u re 3 , le ft p a n e l . I t is,
h o w e ve r, in c re a sin g ly a p p a re n t th a t th e g ro w th d e re g u - o f th e tu m o rs b e a r m u ta n t ras o n c o g e n e s K in zle r a n d
Vo g e lste in , 1 9 9 6 . We su g g e st th a t th e re m a in in g c o lo n ic la tio n w ith in a tu m o r c a n o n ly b e e x p la in e d o n c e w e
u n d e rsta n d th e c o n trib u tio n s o f th e a n c illa ry c e lls p re s- tu m o rs c a rry d e fe c ts in o th e r c o m p o n e n ts o f th e g ro w th
sig n a lin g p a th w a ys th a t p h e n o c o p y ras o n c o g e n e a c ti- e n t in a tu m o r th e a p p a re n tly n o rm a l b ysta n d e rs su c h
a s fib ro b la sts a n d e n d o th e lia l c e lls w h ic h m u st p la y va tio n . T h e n a tu re o f th e se a lte rn a tive , g ro w th -stim u la t-
in g m e c h a n ism s re m a in s e lu sive . k e y ro le s in d rivin g tu m o r c e ll p ro life ra tio n F ig u re 3 ,
rig h t p a n e l . With in n o rm a l tissu e , c e lls a re la rg e ly in - U n d e r in te n sive stu d y fo r tw o d e c a d e s, th e w irin g
d ia g ra m o f th e g ro w th sig n a lin g c irc u itry o f th e m a m m a - stru c te d to g ro w b y th e ir n e ig h b o rs p a ra c rin e sig n a ls
o r via syste m ic e n d o c rin e sig n a ls. C e ll-to -c e ll g ro w th lia n c e ll is c o m in g in to fo c u s F ig u re 2 . N e w d o w n stre a m
e ffe c to r p a th w a ys th a t ra d ia te fro m th e c e n tra l S O S - sig n a lin g is lik e ly to o p e ra te in th e va st m a jo rity o f h u m a n
tu m o rs a s w e ll; virtu a lly a ll a re c o m p o se d o f se ve ra l R a s-R a f-M A P k in a se m ito g e n ic c a sc a d e a re b e in g d is-
c o ve re d w ith so m e re g u la rity H u n te r, 1 9 9 7 ; R o m m e l d istin c t c e ll typ e s th a t a p p e a r to c o m m u n ic a te via h e t-
e ro typ ic sig n a lin g . a n d H a fe n , 1 9 9 8 . T h is c a sc a d e is a lso lin k e d via a va rie ty
o f c ro ss-ta lk in g c o n n e c tio n s w ith o th e r p a th w a ys; th e se H e te ro typ ic sig n a lin g b e tw e e n th e d ive rse c e ll typ e s
w ith in a tu m o r m a y u ltim a te ly p ro ve to b e a s im p o rta n t c ro ss c o n n e c tio n s e n a b le e x tra c e llu la r sig n a ls to e lic it
C e ll
6 0
F ig u re 3 . T u m o rs a s C o m p le x T issu e s
T h e fie ld o f c a n c e r re se a rc h h a s la rg e ly b e e n
g u id e d b y a re d u c tio n ist fo c u s o n c a n c e r c e lls
a n d th e g e n e s w ith in th e m le ft p a n e l a fo -
c u s th a t h a s p ro d u c e d a n e x tra o rd in a ry b o d y
o f k n o w le d g e . L o o k in g fo rw a rd in tim e , w e
b e lie ve th a t im p o rta n t n e w in ro a d s w ill c o m e
fro m re g a rd in g tu m o rs a s c o m p le x tissu e s in
w h ic h m u ta n t c a n c e r c e lls h a ve c o n sc rip te d
a n d su b ve rte d n o rm a l c e ll typ e s to se rve a s
a c tive c o lla b o ra to rs in th e ir n e o p la stic a g e n d a
rig h t p a n e l . T h e in te ra c tio n s b e tw e e n th e
g e n e tic a lly a lte re d m a lig n a n t c e lls a n d th e se
su p p o rtin g c o c o n sp ira to rs w ill p ro ve c ritic a l
to u n d e rsta n d in g c a n c e r p a th o g e n e sis a n d to
th e d e ve lo p m e n t o f n o ve l, e ffe c tive th e ra p ie s.
in e x p la in in g tu m o r c e ll p ro life ra tio n a s th e c a n c e r c e ll- th e c o m p o n e n ts g o ve rn in g th e tra n sit o f th e c e ll th ro u g h
th e G 1 p h a se o f its g ro w th c yc le . C e lls m o n ito r th e ir a u to n o m o u s m e c h a n ism s e n u m e ra te d a b o ve . F o r e x -
a m p le , w e su sp e c t th a t m a n y o f th e g ro w th sig n a ls d riv- e x te rn a l e n viro n m e n t d u rin g th is p e rio d a n d , o n th e b a -
sis o f se n se d sig n a ls, d e c id e w h e th e r to p ro life ra te , to in g th e p ro life ra tio n o f c a rc in o m a c e lls o rig in a te fro m
th e stro m a l c e ll c o m p o n e n ts o f th e tu m o r m a ss. Wh ile b e q u ie sc e n t, o r to e n te r in to a p o stm ito tic sta te . A t th e
m o le c u la r le ve l, m a n y a n d p e rh a p s a ll a n tip ro life ra tive d iffic u lt to va lid a te a t p re se n t, su c h th in k in g re c a sts th e
lo g ic o f a c q u ire d G S a u to n o m y: su c c e ssfu l tu m o r c e lls sig n a ls a re fu n n e le d th ro u g h th e re tin o b la sto m a p ro te in
p R b a n d its tw o re la tive s, p 1 0 7 a n d p 1 3 0 . Wh e n in a a re th o se th a t h a ve a c q u ire d th e a b ility to c o -o p t th e ir
n o rm a l n e ig h b o rs b y in d u c in g th e m to re le a se a b u n d a n t h yp o p h o sp h o ryla te d sta te , p R b b lo c k s p ro life ra tio n b y
se q u e ste rin g a n d a lte rin g th e fu n c tio n o f E 2 F tra n sc rip - flu x e s o f g ro w th -stim u la tin g sig n a ls S k o b e a n d F u -
se n ig , 1 9 9 8 . I n d e e d , in so m e tu m o rs, th e se c o o p e ra tin g tio n fa c to rs th a t c o n tro l th e e x p re ssio n o f b a n k s o f g e n e s
e sse n tia l fo r p ro g re ssio n fro m G 1 in to S p h a se We in - c e lls m a y e ve n tu a lly d e p a rt fro m n o rm a lc y, c o e vo lvin g
w ith th e ir m a lig n a n t n e ig h b o rs in o rd e r to su sta in th e b e rg , 1 9 9 5 .
D isru p tio n o f th e p R b p a th w a y lib e ra te s E 2 F s a n d g ro w th o f th e la tte r K in zle r a n d Vo g e lste in , 1 9 9 8 ; O lu m i
e t a l. , 1 9 9 9 . F u rth e r, in fla m m a to ry c e lls a ttra c te d to site s th u s a llo w s c e ll p ro life ra tio n , re n d e rin g c e lls in se n sitive
to a n tig ro w th fa c to rs th a t n o rm a lly o p e ra te a lo n g th is o f n e o p la sia m a y p ro m o te ra th e r th a n e lim in a te c a n c e r
c e lls C o rd o n -C a rd o a n d P rive s, 1 9 9 9 ; C o u sse n s e t a l. , p a th w a y to b lo c k a d va n c e th ro u g h th e G 1 p h a se o f th e
c e ll c yc le . T h e e ffe c ts o f th e so lu b le sig n a lin g m o le c u le 1 9 9 9 ; H u d so n e t a l. , 1 9 9 9 , a n o th e r e x a m p le o f n o rm a l
c e lls c o n sc rip te d to e n h a n c e tu m o r g ro w th p o te n tia l, T G F a re th e b e st d o c u m e n te d , b u t w e e n visio n o th e r
a n tig ro w th fa c to rs w ill b e fo u n d to sig n a l th ro u g h th is a n o th e r m e a n s to a c q u ire n e c e ssa ry c a p a b ilitie s.
p a th w a y a s w e ll. T G F a c ts in a n u m b e r o f w a ys, m o st
still e lu sive , to p re ve n t th e p h o sp h o ryla tio n th a t in a c ti- Acquired Capability: Insensitivity
to AntigrowthSignals va te s p R b ; in th is fa sh io n , T G F b lo c k s a d va n c e th ro u g h
G 1 . I n so m e c e ll typ e s, T G F su p p re sse s e x p re ssio n With in a n o rm a l tissu e , m u ltip le a n tip ro life ra tive sig n a ls
o p e ra te to m a in ta in c e llu la r q u ie sc e n c e a n d tissu e h o - o f th e c-myc g e n e , w h ic h re g u la te s th e G 1 c e ll c yc le
m a c h in e ry in still u n k n o w n w a ys M o se s e t a l. , 1 9 9 0 . m e o sta sis; th e se sig n a ls in c lu d e b o th so lu b le g ro w th
in h ib ito rs a n d im m o b ilize d in h ib ito rs e m b e d d e d in th e M o re d ire c tly, T G F c a u se s syn th e sis o f th e p 1 5
I N K 4 B
a n d
p 2 1 p ro te in s, w h ic h b lo c k th e c yc lin : C D K c o m p le x e s e x tra c e llu la r m a trix a n d o n th e su rfa c e s o f n e a rb y c e lls.
T h e se g ro w th -in h ib ito ry sig n a ls, lik e th e ir p o sitive ly a c t- re sp o n sib le fo r p R b p h o sp h o ryla tio n H a n n o n a n d
B e a c h , 1 9 9 4 ; D a tto e t a l. , 1 9 9 7 . in g c o u n te rp a rts, a re re c e ive d b y tra n sm e m b ra n e c e ll
su rfa c e re c e p to rs c o u p le d to in tra c e llu la r sig n a lin g c ir- T h e p R b sig n a lin g c irc u it, a s g o ve rn e d b y T G F a n d
o th e r e x trin sic fa c to rs, c a n b e d isru p te d in a va rie ty o f c u its.
A n tig ro w th sig n a ls c a n b lo c k p ro life ra tio n b y tw o d is- w a ys in d iffe re n t typ e s o f h u m a n tu m o rs F yn a n a n d
R e iss, 1 9 9 3 . S o m e lo se T G F re sp o n sive n e ss th ro u g h tin c t m e c h a n ism s. C e lls m a y b e fo rc e d o u t o f th e a c tive
p ro life ra tive c yc le in to th e q u ie sc e n t G
0
sta te fro m d o w n re g u la tio n o f th e ir T G F re c e p to rs, w h ile o th e rs
d isp la y m u ta n t, d ysfu n c tio n a l re c e p to rs F yn a n a n d w h ic h th e y m a y re e m e rg e o n so m e fu tu re o c c a sio n
w h e n e x tra c e llu la r sig n a ls p e rm it. A lte rn a tive ly, c e lls R e iss, 1 9 9 3 ; M a rk o w itz e t a l. , 1 9 9 5 . T h e c yto p la sm ic
S m a d 4 p ro te in , w h ic h tra n sd u c e s sig n a ls fro m lig a n d - m a y b e in d u c e d to p e rm a n e n tly re lin q u ish th e irp ro life ra -
tive p o te n tia l b y b e in g in d u c e d to e n te r in to p o stm ito tic a c tiva te d T G F re c e p to rs to d o w n stre a m ta rg e ts, m a y
b e e lim in a te d th ro u g h m u ta tio n o f its e n c o d in g g e n e sta te s, u su a lly a sso c ia te d w ith a c q u isitio n o f sp e c ific
d iffe re n tia tio n -a sso c ia te d tra its. S c h u tte e t a l. , 1 9 9 6 . T h e lo c u s e n c o d in g p 1 5
I N K 4 B
m a y b e
d e le te d C h in e t a l. , 1 9 9 8 . A lte rn a tive ly, th e im m e d ia te I n c ip ie n t c a n c e r c e lls m u st e va d e th e se a n tip ro life ra -
tive sig n a ls if th e y a re to p ro sp e r. M u c h o f th e c irc u itry d o w n stre a m ta rg e t o f its a c tio n s, C D K 4 , m a y b e c o m e
u n re sp o n sive to th e in h ib ito ry a c tio n s o f p 1 5
I N K 4 B
b e - th a t e n a b le s n o rm a l c e lls to re sp o n d to a n tig ro w th sig -
n a ls is a sso c ia te d w ith th e c e ll c yc le c lo c k , sp e c ific a lly c a u se o f m u ta tio n s th a t c re a te a m in o a c id su b stitu tio n s
R e vie w
6 1
in its I N K 4 A /B -in te ra c tin g d o m a in ; th e re su ltin g c yc lin in virtu a lly a ll c e ll typ e s th ro u g h o u t th e b o d y. O n c e trig -
g e re d b y a va rie ty o f p h ysio lo g ic sig n a ls, th is p ro g ra m D : C D K 4 c o m p le x e s a re th e n g ive n a fre e h a n d to in a c ti-
va te p R b b y h yp e rp h o sp h o ryla tio n Z u o e t a l. , 1 9 9 6 . u n fo ld s in a p re c ise ly c h o re o g ra p h e d se rie s o f ste p s.
C e llu la r m e m b ra n e s a re d isru p te d , th e c yto p la sm ic a n d F in a lly, fu n c tio n a l p R b , th e e n d ta rg e t o f th is p a th w a y,
m a y b e lo st th ro u g h m u ta tio n o f its g e n e . A lte rn a tive ly, n u c le a r sk e le to n s a re b ro k e n d o w n , th e c yto so l is e x -
tru d e d , th e c h ro m o so m e s a re d e g ra d e d , a n d th e n u - in c e rta in D N A viru s-in d u c e d tu m o rs, n o ta b ly c e rvic a l
c a rc in o m a s, p R b fu n c tio n is e lim in a te d th ro u g h se q u e s- c le u s is fra g m e n te d , a ll in a sp a n o f 3 0 1 2 0 m in . I n th e
e n d , th e sh rive le d c e ll c o rp se is e n g u lfe d b y n e a rb y c e lls tra tio n b y vira l o n c o p ro te in s, su c h a s th e E 7 o n c o p ro te in
o f h u m a n p a p illo m a viru s D yso n e t a l. , 1 9 8 9 . I n a d d itio n , in a tissu e a n d d isa p p e a rs, typ ic a lly w ith in 2 4 h r Wyllie
e t a l. , 1 9 8 0 . c a n c e r c e lls c a n a lso tu rn o ff e xp re ssio n o f in te g rin s a n d
o th e r c e ll a d h e sio n m o le c u le s th a t se n d a n tig ro w th sig - T h e a p o p to tic m a c h in e ry c a n b e b ro a d ly d ivid e d in to
tw o c la sse s o f c o m p o n e n ts se n so rs a n d e ffe c to rs. T h e n a ls, fa vo rin g in ste a d th o se th a t c o n ve y p ro g ro w th sig -
n a ls; th e se a d h e re n c e -b a se d a n tig ro w th sig n a ls lik e ly se n so rs a re re sp o n sib le fo r m o n ito rin g th e e x tra c e llu la r
a n d in tra c e llu la r e n viro n m e n t fo r c o n d itio n s o f n o rm a lity im p in g e o n th e p R b c irc u it a s w e ll. T h e b o tto m lin e is
th a t th e a n tig ro w th c irc u it c o n ve rg in g o n to R b a n d th e o r a b n o rm a lity th a t in flu e n c e w h e th e r a c e ll sh o u ld live
o r d ie . T h e se sig n a ls re g u la te th e se c o n d c la ss o f c o m - c e ll d ivisio n c yc le is, o n e w a y o r a n o th e r, d isru p te d in
a m a jo rity o f h u m a n c a n c e rs, d e fin in g th e c o n c e p t a n d p o n e n ts, w h ic h fu n c tio n a s e ffe c to rs o f a p o p to tic d e a th .
T h e se n tin e ls in c lu d e c e ll su rfa c e re c e p to rs th a t b in d a p u rp o se o f tu m o r su p p re sso r lo ss in c a n c e r.
C e ll p ro life ra tio n d e p e n d s o n m o re th a n a n a vo id a n c e su rviva l o r d e a th fa c to rs. E x a m p le s o f th e se lig a n d /
re c e p to r p a irs in c lu d e su rviva l sig n a ls c o n ve ye d b y I G F - o f c yto sta tic a n tig ro w th sig n a ls. O u r tissu e s a lso c o n -
stra in c e ll m u ltip lic a tio n b y in stru c tin g c e lls to e n te r irre - 1 /I G F -2 th ro u g h th e ir re c e p to r, I G F -1 R , a n d b y I L -3 a n d
its c o g n a te re c e p to r, I L -3 R L o te m a n d S a c h s, 1 9 9 6 ; ve rsib ly in to p o stm ito tic , d iffe re n tia te d sta te s, u sin g d i-
ve rse m e c h a n ism s th a t a re in c o m p le te ly u n d e rsto o d ; it B u tt e t a l. , 1 9 9 9 . D e a th sig n a ls a re c o n ve ye d b y th e
F A S lig a n d b in d in g th e F A S re c e p to r a n d b y T N F b in d - is a p p a re n t th a t tu m o r c e lls u se va rio u s stra te g ie s to
a vo id th is te rm in a l d iffe re n tia tio n . O n e stra te g y fo r in g T N F -R 1 A sh k e n a zi a n d D ix it, 1 9 9 9 . I n tra c e llu la r
se n so rs m o n ito r th e c e lls w e ll-b e in g a n d a c tiva te th e a vo id in g d iffe re n tia tio n d ire c tly in vo lve s th e c-myc o n -
c o g e n e , w h ic h e n c o d e s a tra n sc rip tio n fa c to r. D u rin g d e a th p a th w a y in re sp o n se to d e te c tin g a b n o rm a litie s,
in c lu d in g D N A d a m a g e , sig n a lin g im b a la n c e p ro vo k e d n o rm a l d e ve lo p m e n t, th e g ro w th -stim u la tin g a c tio n o f
M yc , in a sso c ia tio n w ith a n o th e r fa c to r, M a x , c a n b e b y o n c o g e n e a c tio n , su rviva l fa c to r in su ffic ie n c y, o r h yp -
o x ia E va n a n d L ittle w o o d , 1 9 9 8 . F u rth e r, th e life o f m o st su p p la n te d b y a lte rn a tive c o m p le x e s o f M a x w ith a
g ro u p o f M a d tra n sc rip tio n fa c to rs; th e M a d M a x c o m - c e lls is in p a rt m a in ta in e d b y c e llm a trix a n d c e llc e ll
a d h e re n c e -b a se d su rviva l sig n a ls w h o se a b ro g a tio n p le x e s e lic it d iffe re n tia tio n -in d u c in g sig n a ls F o le y a n d
E ise n m a n , 1 9 9 9 . H o w e ve r, o ve re x p re ssio n o f th e c -M yc e lic its a p o p to sis I sh iza k i e t a l. , 1 9 9 5 ; G ia n c o tti a n d R u -
o sla h ti, 1 9 9 9 . B o th so lu b le a n d im m o b ilize d a p o p to tic o n c o p ro te in , a s is se e n in m a n y tu m o rs, c a n re ve rse th is
p ro c e ss, sh iftin g th e b a la n c e b a c k to fa vo r M yc M a x re g u la to ry sig n a ls lik e ly re fle c t th e n e e d s o f tissu e s to
m a in ta in th e ir c o n stitu e n t c e lls in a p p ro p ria te a rc h ite c - c o m p le x e s, th e re b y im p a irin g d iffe re n tia tio n a n d p ro -
m o tin g g ro w th . D u rin g h u m a n c o lo n c a rc in o g e n e sis, in - tu ra l c o n fig u ra tio n s.
M a n y o f th e sig n a ls th a t e lic it a p o p to sis c o n ve rg e a c tiva tio n o f th e A P C /-c a te n in p a th w a y se rve s to b lo c k
th e e g re ss o f e n te ro c yte s in th e c o lo n ic c ryp ts in to a o n th e m ito c h o n d ria , w h ic h re sp o n d to p ro a p o p to tic
sig n a ls b y re le a sin g c yto c h ro m e C , a p o te n t c a ta lyst o f d iffe re n tia te d , p o stm ito tic sta te K in zle r a n d Vo g e lste in ,
1 9 9 6 . A n a lo g o u sly, d u rin g th e g e n e ra tio n o f a via n e ryth - a p o p to sis G re e n a n d R e e d , 1 9 9 8 . M e m b e rs o f th e B c l-2
fa m ily o f p ro te in s, w h o se m e m b e rs h a ve e ith e r p ro - ro b la sto sis, th e erbA o n c o g e n e a c ts to p re ve n t irre ve rs-
ib le e ryth ro c yte d iffe re n tia tio n K a h n e t a l. , 1 9 8 6 . a p o p to tic B a x , B a k , B id , B im o r a n tia p o p to tic B c l-2 ,
B c l-XL , B c l-W fu n c tio n , a c t in p a rt b y g o ve rn in g m ito - Wh ile th e c o m p o n e n ts a n d in te rc o n n e c tio n s b e tw e e n
th e va rio u s a n tig ro w th a n d d iffe re n tia tio n -in d u c in g sig - c h o n d ria l d e a th sig n a lin g th ro u g h c yto c h ro m e C re -
le a se . T h e p 5 3 tu m o r su p p re sso r p ro te in c a n e lic it a p o - n a ls a n d th e c o re c e ll c yc le m a c h in e ry a re still b e in g
d e lin e a te d , th e e x iste n c e o f a n a n tig ro w th sig n a lin g c ir- p to sis b y u p re g u la tin g e x p re ssio n o f p ro a p o p to tic B a x
in re sp o n se to se n sin g D N A d a m a g e ; B a x in tu rn stim u - c u itry is c le a r F ig u re 2 , a s is th e n e c e ssity fo r its c irc u m -
ve n tio n b y d e ve lo p in g c a n c e rs. la te s m ito c h o n d ria to re le a se c yto c h ro m e C .
T h e u ltim a te e ffe c to rs o f a p o p to sis in c lu d e a n a rra y
o f in tra c e llu la r p ro te a se s te rm e d c a sp a se s T h o rn b e rry
Acquired Capability: Evading Apoptosis
a n d L a ze b n ik , 1 9 9 8 . T w o g a te k e e p e r c a sp a se s, 8
T h e a b ility o f tu m o r c e ll p o p u la tio n s to e x p a n d in n u m b e r
a n d 9 , a re a c tiva te d b y d e a th re c e p to rs su c h a s F A S is d e te rm in e d n o t o n ly b y th e ra te o f c e ll p ro life ra tio n
o r b y th e c yto c h ro m e C re le a se d fro m m ito c h o n d ria , b u t a lso b y th e ra te o f c e ll a ttritio n . P ro g ra m m e d c e ll
re sp e c tive ly. T h e se p ro x im a l c a sp a se s trig g e r th e a c ti- d e a th a p o p to sis re p re se n ts a m a jo r so u rc e o f th is
va tio n o f a d o ze n o rm o re e ffe c to r c a sp a se s th a t e x e c u te a ttritio n . T h e e vid e n c e is m o u n tin g , p rin c ip a lly fro m
th e d e a th p ro g ra m , th ro u g h se le c tive d e stru c tio n o f su b - stu d ie s in m o u se m o d e ls a n d c u ltu re d c e lls, a s w e ll a s
c e llu la r stru c tu re s a n d o rg a n e lle s, a n d o f th e g e n o m e . fro m d e sc rip tive a n a lyse s o f b io p sie d sta g e s in h u m a n
T h e p o ssib ility th a t a p o p to sis se rve s a s a b a rrie r to c a rc in o g e n e sis, th a t a c q u ire d re sista n c e to w a rd a p o -
c a n c e r w a s first ra ise d in 1 9 7 2 , w h e n K e rr, Wyllie , a n d p to sis is a h a llm a rk o f m o st a n d p e rh a p s a ll typ e s o f
C u rrie d e sc rib e d m a ssive a p o p to sis in th e c e lls p o p u la t- c a n c e r.
in g ra p id ly g ro w in g , h o rm o n e -d e p e n d e n t tu m o rs fo llo w - O b se rva tio n s a c c u m u la te d o ve r th e p a st d e c a d e in d i-
c a te th a t th e a p o p to tic p ro g ra m is p re se n t in la te n t fo rm in g h o rm o n e w ith d ra w a l K e rr e t a l. , 1 9 7 2 . T h e d isc o ve ry
C e ll
6 2
o f th e bcl-2 o n c o g e n e b y its u p re g u la tio n via c h ro m o - a b n o rm a litie s, in c lu d in g h yp o x ia a n d o n c o g e n e h yp e r-
e x p re ssio n , a re a lso fu n n e le d in p a rt via p 5 3 to th e a p o - so m a l tra n slo c a tio n in fo llic u la r lym p h o m a re vie w e d in
p to tic m a c h in e ry; th e se to o a re im p a ire d a t e lic itin g K o rsm e ye r, 1 9 9 2 a n d its re c o g n itio n a s h a vin g a n ti-
a p o p to sis w h e n p 5 3 fu n c tio n is lo st L e vin e , 1 9 9 7 . A d d i- a p o p to tic a c tivity Va u x e t a l. , 1 9 8 8 o p e n e d u p th e in -
tio n a lly, th e P I 3 k in a se A K T /P K B p a th w a y, w h ic h tra n s- ve stig a tio n o f a p o p to sis in c a n c e r a t th e m o le c u la r le ve l.
m its a n tia p o p to tic su rviva l sig n a ls, is lik e ly in vo lve d in Wh e n c o e x p re sse d w ith a myc o n c o g e n e in tra n sg e n ic
m itig a tin g a p o p to sis in a su b sta n tia l fra c tio n o f h u m a n m ic e , th e bcl-2 g e n e w a s a b le to p ro m o te fo rm a tio n o f
tu m o rs. T h is su rviva l sig n a lin g c irc u it c a n b e a c tiva te d
B c e ll lym p h o m a s b y e n h a n c in g lym p h o c yte su rviva l, n o t
b y e x tra c e llu la r fa c to rs su c h a s I G F -1 /2 o r I L -3 E va n
b y fu rth e r stim u la tin g th e ir myc-in d u c e d p ro life ra tio n
a n d L ittle w o o d , 1 9 9 8 , b y in tra c e llu la r sig n a ls e m a n a tin g
S tra sse r e t a l. , 1 9 9 0 ; fu rth e r, 5 0 % o f th e in fre q u e n t
fro m R a s D o w n w a rd , 1 9 9 8 , o r b y lo ss o f th e p T E N
lym p h o m a s a risin g in bcl-2sin g le tra n sg e n ic tra n sg e n ic
tu m o r su p p re sso r, a p h o sp h o lip id p h o sp h a ta se th a t
m ic e h a d so m a tic tra n slo c a tio n s a c tiva tin g c-myc, c o n -
n o rm a lly a tte n u a te s th e A K T su rviva l sig n a l C a n tle y a n d
firm in g a se le c tive p re ssu re d u rin g lym p h o m a g e n e sis
N e e l, 1 9 9 9 . R e c e n tly, a m e c h a n ism fo r a b ro g a tin g th e
to u p re g u la te b o th B c l-2 a n d c -M yc M c D o n n e ll a n d
F A S d e a th sig n a l h a s b e e n re ve a le d in a h ig h fra c tio n
K o rsm e ye r, 1 9 9 1 .
o f lu n g a n d c o lo n c a rc in o m a c e ll lin e s: a n o n sig n a lin g
F u rth e r in sig h t in to th e myc-bcl-2in te ra c tio n e m e rg e d
d e c o y re c e p to r fo r F A S lig a n d is u p re g u la te d , titra tin g
la te r fro m stu d yin g th e e ffe c ts o f a myc o n c o g e n e o n
th e d e a th -in d u c in g sig n a l a w a y fro m th e F A S d e a th re -
fib ro b la sts c u ltu re d in lo w se ru m . Wid e sp re a d a p o p to sis
c e p to r P itti e t a l. , 1 9 9 8 . We e x p e c t th a t virtu a lly a ll
w a s in d u c e d in myc-e x p re ssin g c e lls la c k in g se ru m ; th e
c a n c e r c e lls h a rb o r a lte ra tio n s th a t e n a b le e va sio n o f
c o n se q u e n t a p o p to sis c o u ld b e a b ro g a te d b y e x o g e -
a p o p to sis.
n o u s su rviva l fa c to rs e . g . , I G F -1 , b y fo rc e d o ve re x p re s-
I t is n o w p o ssib le to la y o u t a p ro visio n a l a p o p to tic
sio n o f B c l-2 o r th e re la te d B c l-XL p ro te in , o r b y d isru p -
sig n a lin g c irc u itry F ig u re 2 ; w h ile in c o m p le te , it is e vi-
tio n o f th e F A S d e a th sig n a lin g c irc u it H u e b e r e t a l. ,
d e n t th a t m o st re g u la to ry a n d e ffe c to r c o m p o n e n ts a re
1 9 9 7 . C o lle c tive ly, th e d a ta in d ic a te th a t a c e lls a p o -
p re se n t in re d u n d a n t fo rm . T h is re d u n d a n c y h o ld s im -
p to tic p ro g ra m c a n b e trig g e re d b y a n o ve re x p re sse d
p o rta n t im p lic a tio n s fo r th e d e ve lo p m e n t o f n o ve l typ e s
o n c o g e n e . I n d e e d , e lim in a tio n o f c e lls b e a rin g a c tiva te d
o f a n titu m o r th e ra p y, sin c e tu m o r c e lls th a t h a ve lo st
o n c o g e n e s b y a p o p to sis m a y re p re se n t th e p rim a ry
p ro a p o p to tic c o m p o n e n ts a re lik e ly to re ta in o th e r sim i-
m e a n s b y w h ic h su c h m u ta n t c e lls a re c o n tin u a lly c u lle d
la r o n e s. We a n tic ip a te th a t n e w te c h n o lo g ie s w ill b e
fro m th e b o d ys tissu e s.
a b le to d isp la y th e a p o p to tic p a th w a ys still o p e ra tive in
O th e r e x a m p le s stre n g th e n th e c o n se n su s th a t a p o -
sp e c ific typ e s o f c a n c e r c e lls a n d th a t n e w d ru g s w ill
p to sis is a m a jo r b a rrie r to c a n c e r th a t m u st b e c irc u m -
e n a b le c ro ss-ta lk b e tw e e n th e still in ta c t c o m p o n e n ts
ve n te d . T h u s, in tra n sg e n ic m ic e w h e re th e p R b tu m o r
o f p a ra lle l a p o p to tic sig n a lin g p a th w a ys in tu m o r c e lls,
su p p re sso r w a s fu n c tio n a lly in a c tiva te d in th e c h o ro id
re su ltin g in re sto ra tio n o f th e a p o p to tic d e fe n se m e c h a -
p le x u s, slo w ly g ro w in g m ic ro sc o p ic tu m o rs a ro se , e x -
n ism , w ith su b sta n tia l th e ra p e u tic b e n e fit.
h ib itin g h ig h a p o p to tic ra te s; th e a d d itio n a l in a c tiva tio n
o f th e p 5 3 tu m o r su p p re sso r p ro te in , a c o m p o n e n t o f
Acquired Capability: Limitless Replicative Potential
th e a p o p to tic sig n a lin g c irc u itry, le d to ra p id ly g ro w in g
T h re e a c q u ire d c a p a b ilitie s g ro w th sig n a l a u to n o m y,
tu m o rs c o n ta in in g lo w n u m b e rs o f a p o p to tic c e lls S y-
in se n sitivity to a n tig ro w th sig n a ls, a n d re sista n c e to
m o n d s e t a l. , 1 9 9 4 . T h e ro le o f e x tra c e llu la r su rviva l
a p o p to sis a ll le a d to a n u n c o u p lin g o f a c e lls g ro w th
fa c to rs is illu stra te d b y d ise a se p ro g re ssio n in tra n s-
p ro g ra m fro m sig n a ls in its e n viro n m e n t. I n p rin c ip le ,
g e n ic m ic e p ro n e to p a n c re a tic isle t tu m o rs. I f IGF-2
th e re su ltin g d e re g u la te d p ro life ra tio n p ro g ra m sh o u ld
g e n e e x p re ssio n , w h ic h is a c tiva te d in th is tu m o rig e n e -
su ffic e to e n a b le th e g e n e ra tio n o f th e va st c e ll p o p u la -
sis p a th w a y, w a s a b ro g a te d u sin g g e n e k n o c k o u t m ic e ,
tio n s th a t c o n stitu te m a c ro sc o p ic tu m o rs. H o w e ve r, re -
tu m o r g ro w th a n d p ro g re ssio n w e re im p a ire d , a s e vi-
se a rc h p e rfo rm e d o ve r th e p a st 3 0 ye a rs in d ic a te s th a t
d e n c e d b y th e a p p e a ra n c e o f c o m p a ra tive ly sm a ll, b e -
th is a c q u ire d d isru p tio n o f c e ll-to -c e ll sig n a lin g , o n its
n ig n tu m o rs sh o w in g h ig h ra te s o f a p o p to sis C h risto fo ri
o w n , d o e s n o t e n su re e x p a n sive tu m o r g ro w th . M a n y
e t a l. , 1 9 9 4 . I n th e se c e lls, th e a b se n c e o f I G F -2 d id n o t
a n d p e rh a p s a ll typ e s o f m a m m a lia n c e lls c a rry a n in trin -
a ffe c t c e ll p ro life ra tio n ra te s, c le a rly id e n tifyin g it a s a n
sic , c e ll-a u to n o m o u s p ro g ra m th a t lim its th e irm u ltip lic a -
a n tia p o p to tic su rviva l fa c to r. C o lle c tive ly, th e se o b se r-
tio n . T h is p ro g ra m a p p e a rs to o p e ra te in d e p e n d e n tly o f
va tio n s a rg u e th a t a lte rin g c o m p o n e n ts o f th e a p o p to tic
th e c e ll-to -c e ll sig n a lin g p a th w a ys d e sc rib e d a b o ve . I t
m a c h in e ry c a n d ra m a tic a lly a ffe c t th e d yn a m ic s o f tu -
to o m u st b e d isru p te d in o rd e r fo r a c lo n e o f c e lls to
m o r p ro g re ssio n , p ro vid in g a ra tio n a le fo r th e in a c tiva -
e x p a n d to a size th a t c o n stitu te s a m a c ro sc o p ic , life -
tio n o f th is m a c h in e ry d u rin g tu m o r d e ve lo p m e n t.
th re a te n in g tu m o r.
R e sista n c e to a p o p to sis c a n b e a c q u ire d b y c a n c e r
T h e e a rly w o rk o f H a yflic k d e m o n stra te d th a t c e lls in
c e lls th ro u g h a va rie ty o f stra te g ie s. S u re ly, th e m o st
c u ltu re h a ve a fin ite re p lic a tive p o te n tia l re vie w e d in
c o m m o n ly o c c u rrin g lo ss o f a p ro a p o p to tic re g u la to r
H a yflic k , 1 9 9 7 . O n c e su c h c e ll p o p u la tio n s h a ve p ro -
th ro u g h m u ta tio n in vo lve s th e p53 tu m o r su p p re sso r g re sse d th ro u g h a c e rta in n u m b e r o f d o u b lin g s, th e y
g e n e . T h e re su ltin g fu n c tio n a l in a c tiva tio n o f its p ro d u c t, sto p g ro w in g a p ro c e ss te rm e d se n e sc e n c e . T h e se -
th e p 5 3 p ro te in , is se e n in g re a te r th a n 5 0 % o f h u m a n n e sc e n c e o f c u ltu re d h u m a n fib ro b la sts c a n b e c irc u m -
c a n c e rs a n d re su lts in th e re m o va l o f a k e y c o m p o n e n t ve n te d b y d isa b lin g th e ir p R b a n d p 5 3 tu m o r su p p re sso r
o f th e D N A d a m a g e se n so r th a t c a n in d u c e th e a p o p to tic p ro te in s, e n a b lin g th e se c e lls to c o n tin u e m u ltip lyin g fo r
a d d itio n a l g e n e ra tio n s u n til th e y e n te r in to a se c o n d e ffe c to r c a sc a d e H a rris, 1 9 9 6 . S ig n a ls e vo k e d b y o th e r
R e vie w
6 3
sta te te rm e d c risis. T h e c risis sta te is c h a ra c te rize d b y th re sh o ld , a n d th is in tu rn p e rm its u n lim ite d m u ltip lic a -
m a ssive c e ll d e a th , k a ryo typ ic d isa rra y a sso c ia te d w ith tio n o f d e sc e n d a n t c e lls. B o th m e c h a n ism s se e m to b e
e n d -to -e n d fu sio n o f c h ro m o so m e s, a n d th e o c c a sio n a l stro n g ly su p p re sse d in m o st n o rm a l h u m a n c e lls in o rd e r
e m e rg e n c e o f a va ria n t 1 in 1 0
7
c e ll th a t h a s a c q u ire d to d e n y th e m u n lim ite d re p lic a tive p o te n tia l.
th e a b ility to m u ltip ly w ith o u t lim it, th e tra it te rm e d im - T h e ro le o f te lo m e ra se in im m o rta lizin g c e lls c a n b e
m o rta liza tio n Wrig h t e t a l. , 1 9 8 9 . d e m o n stra te d d ire c tly b y e c to p ic a lly e x p re ssin g th e e n -
P ro vo c a tive ly, m o st typ e s o f tu m o r c e lls th a t a re p ro p - zym e in c e lls, w h e re it c a n c o n ve y u n lim ite d re p lic a tive
a g a te d in c u ltu re a p p e a r to b e im m o rta lize d , su g g e stin g p o te n tia l o n to a va rie ty o f n o rm a l e a rly p a ssa g e , p re se -
th a t lim itle ss re p lic a tive p o te n tia l is a p h e n o typ e th a t n e sc e n t c e lls in vitro B o d n a r e t a l. , 1 9 9 8 ; Va ziri a n d
w a s a c q u ire d in vivo d u rin g tu m o r p ro g re ssio n a n d w a s B e n c h im o l, 1 9 9 8 . F u rth e r, la te p a ssa g e c e lls p o ise d to
e sse n tia l fo r th e d e ve lo p m e n t o f th e ir m a lig n a n t g ro w th e n te r c risis c o n tin u e to p ro life ra te w ith o u t g ivin g a n y
sta te H a yflic k , 1 9 9 7 . T h is re su lt su g g e sts th a t a t so m e
e vid e n c e o f c risis w h e n su p p lie d w ith th is e n zym e
p o in t d u rin g th e c o u rse o f m u ltiste p tu m o r p ro g re ssio n ,
C o u n te r e t a l. , 1 9 9 8 ; H a lvo rse n e t a l. , 1 9 9 9 ; Z h u e t a l. ,
e vo lvin g p re m a lig n a n t c e ll p o p u la tio n s e x h a u st th e ir e n -
1 9 9 9 . A d d itio n a l c lu e s in to th e im p o rta n c e o f te lo m e re
d o w m e n t o f a llo w e d d o u b lin g s a n d c a n o n ly c o m p le te
m a in te n a n c e fo r c a n c e r c o m e s fro m a n a lysis o f m ic e
th e ir tu m o rig e n ic a g e n d a b y b re a c h in g th e m o rta lity b a r-
la c k in g te lo m e ra se fu n c tio n . F o r e x a m p le , m ic e c a rryin g
rie r a n d a c q u irin g u n lim ite d re p lic a tive p o te n tia l.
a h o m o zyg o u s k n o c k o u t o f th e c e ll c yc le in h ib ito r
O b se rva tio n s o f c u ltu re d c e lls in d ic a te th a t va rio u s
p 1 6
I N K 4 A
a re tu m o r p ro n e , p a rtic u la rly w h e n e x p o se d to
n o rm a l h u m a n c e ll typ e s h a ve th e c a p a c ity fo r 6 0 7 0
c a rc in o g e n s; th e tu m o rs th a t a rise sh o w c o m p a ra tive ly
d o u b lin g s. T a k e n a t fa c e va lu e , th e se n u m b e rs m a k e
e le va te d te lo m e ra se a c tivity. Wh e n c a rc in o g e n s w e re
little se n se w h e n a tte m p tin g to in vo k e c e ll m o rta lity a s
a p p lie d to p 1 6
I N K 4 A
-n u ll m ic e th a t a lso la c k e d te lo m e ra se ,
a n im p e d im e n t to c a n c e r fo rm a tio n : 6 0 7 0 d o u b lin g s
tu m o r in c id e n c e w a s re d u c e d , c o n c o m ita n t w ith su b -
sh o u ld e n a b le c lo n e s o f tu m o r c e lls to e x p a n d to n u m -
sta n tia l te lo m e re sh o rte n in g a n d k a ryo typ ic d isa rra y in
b e rs th a t va stly e x c e e d th e n u m b e r o f c e lls in th e h u m a n
th o se tu m o rs th a t d id a p p e a r G re e n b e rg e t a l. , 1 9 9 9 .
b o d y. I f c lu e s fro m e va lu a tio n o f p ro life ra tio n a n d a p o -
Wh ile te lo m e re m a in te n a n c e is c le a rly a k e y c o m p o -
p to tic ra te s in c e rta in h u m a n tu m o rs Wyllie e t a l. , 1 9 8 0
n e n t o f th e c a p a b ility fo r u n lim ite d re p lic a tio n , w e re m a in
a n d tra n sg e n ic m o u se m o d e ls S ym o n d s e t a l. , 1 9 9 4 ;
u n c e rta in a b o u t a n o th e r o n e , th e c irc u m ve n tio n o f c e llu -
S h ib a ta e t a l. , 1 9 9 6 ; B e rg e rs e t a l. , 1 9 9 8 p ro ve g e n e ra liz-
la r se n e sc e n c e . T h e p h e n o m e n o n o f se n e sc e n c e w a s
a b le , th e p a ra d o x c a n b e re so lve d : e vo lvin g p re m a lig -
o rig in a lly o b se rve d a s a d e la ye d re sp o n se o f p rim a ry
n a n t a n d m a lig n a n t c e ll p o p u la tio n s e vid e n c e c h ro n ic ,
c e lls to e x te n d e d p ro p a g a tio n in vitro a n d h a s th u s b e e n
w id e sp re a d a p o p to sis a n d c o n se q u e n tly su ffe r c o n sid -
a sso c ia te d w ith m e c h a n ism s o f d ivisio n a l c o u n tin g
e ra b le c e ll a ttritio n c o n c o m ita n t w ith c e ll a c c u m u la tio n .
H a yflic k , 1 9 9 7 . M o re re c e n tly, th e se n e sc e n t sta te h a s
T h u s, th e n u m b e r o f c e lls in a tu m o r g re a tly u n d e rre p re -
b e e n o b se rve d to b e in d u c ib le in c e rta in c u ltu re d c e lls
se n ts th e c e ll g e n e ra tio n s re q u ire d to p ro d u c e it, ra isin g
in re sp o n se to h ig h le ve l e x p re ssio n o f g e n e s su c h a s
th e g e n e ra tio n a l lim it o f n o rm a l so m a tic c e lls a s a b a rrie r
th e a c tiva te d ras o n c o g e n e S e rra n o e t a l. , 1 9 9 7 .
to c a n c e r.
T h e a b o ve -c ite d o b se rva tio n s m ig h t a rg u e th a t se n e s-
T h e c o u n tin g d e vic e fo r c e ll g e n e ra tio n s h a s b e e n
c e n c e , m u c h lik e a p o p to sis, re fle c ts a p ro te c tive m e c h a -
d isc o ve re d o ve r th e p a st d e c a d e : th e e n d s o f c h ro m o -
n ism th a t c a n b e a c tiva te d b y sh o rte n e d te lo m e re s o r
so m e s, te lo m e re s, w h ic h a re c o m p o se d o f se ve ra l th o u -
c o n flic tin g g ro w th sig n a ls th a t fo rc e s a b e rra n t c e lls irre -
sa n d re p e a ts o f a sh o rt 6 b p se q u e n c e e le m e n t. R e p lic a -
ve rsib ly in to a G
0
-lik e sta te , th e re b y re n d e rin g th e m in c a -
tive g e n e ra tio n s a re c o u n te d b y th e 5 0 1 0 0 b p lo ss o f
p a b le o f fu rth e r p ro life ra tio n . I f so , c irc u m ve n tio n o f se -
te lo m e ric D N A fro m th e e n d s o f e ve ry c h ro m o so m e d u r-
n e sc e n c e in vivo m a y in d e e d re p re se n t a n e sse n tia l ste p
in g e a c h c e ll c yc le . T h is p ro g re ssive sh o rte n in g h a s
in tu m o r p ro g re ssio n th a t is re q u ire d fo r th e su b se q u e n t
b e e n a ttrib u te d to th e in a b ility o f D N A p o lym e ra se s to
a p p ro a c h to a n d b re a c h in g o f th e c risis b a rrie r. B u t w e
c o m p le te ly re p lic a te th e 3 e n d s o f c h ro m o so m a l D N A
c o n sid e r a n a lte rn a tive m o d e l e q u a lly p la u sib le : se n e s-
d u rin g e a c h S p h a se . T h e p ro g re ssive e ro sio n o f te lo -
c e n c e c o u ld b e a n a rtifa c t o f c e ll c u ltu re th a t d o e s n o t
m e re s th ro u g h su c c e ssive c yc le s o f re p lic a tio n e ve n tu -
re fle c t a p h e n o typ e o f c e lls w ith in livin g tissu e s a n d
a lly c a u se s th e m to lo se th e ir a b ility to p ro te c t th e e n d s
d o e s n o t re p re se n t a n im p e d im e n t to tu m o r p ro g re ssio n
o f c h ro m o so m a l D N A . T h e u n p ro te c te d c h ro m o so m a l
in vivo . R e so lu tio n o f th is q u a n d a ry w ill b e c ritic a l to
e n d s p a rtic ip a te in e n d -to -e n d c h ro m o so m a l fu sio n s,
c o m p le te ly u n d e rsta n d th e a c q u isitio n o f lim itle ss re p li-
yie ld in g th e k a ryo typ ic d isa rra y a sso c ia te d w ith c risis
c a tive p o te n tia l.
a n d re su ltin g , a lm o st in e vita b ly, in th e d e a th o f th e a f-
fe c te d c e ll C o u n te r e t a l. , 1 9 9 2 .
Acquired Capability: Sustained Angiogenesis
T e lo m e re m a in te n a n c e is e vid e n t in virtu a lly a ll typ e s o f
T h e o x yg e n a n d n u trie n ts su p p lie d b y th e va sc u la tu re
m a lig n a n t c e lls S h a y a n d B a c c h e tti, 1 9 9 7 ; 8 5 % 9 0 %
a re c ru c ia l fo r c e ll fu n c tio n a n d su rviva l, o b lig a tin g virtu -
o f th e m su c c e e d in d o in g so b y u p re g u la tin g e x p re ssio n
a lly a ll c e lls in a tissu e to re sid e w ith in 1 0 0 m o f a
o f th e te lo m e ra se e n zym e , w h ic h a d d s h e x a n u c le o tid e
c a p illa ry b lo o d ve sse l. D u rin g o rg a n o g e n e sis, th is c lo se -
re p e a ts o n to th e e n d s o f te lo m e ric D N A B rya n a n d
n e ss is e n su re d b y c o o rd in a te d g ro w th o f ve sse ls a n d C e c h , 1 9 9 9 , w h ile th e re m a in d e r h a ve in ve n te d a w a y
p a re n c h ym a . O n c e a tissu e is fo rm e d , th e g ro w th o f o f a c tiva tin g a m e c h a n ism , te rm e d A L T , w h ic h a p p e a rs
n e w b lo o d ve sse ls th e p ro c e ss o f a n g io g e n e sis is to m a in ta in te lo m e re s th ro u g h re c o m b in a tio n -b a se d in -
tra n sito ry a n d c a re fu lly re g u la te d . B e c a u se o f th is d e - te rc h ro m o so m a l e x c h a n g e s o f se q u e n c e in fo rm a tio n
p e n d e n c e o n n e a rb y c a p illa rie s, it w o u ld se e m p la u sib le B rya n e t a l. , 1 9 9 5 . B y o n e o r th e o th e r m e c h a n ism ,
te lo m e re s a re m a in ta in e d a t a le n g th a b o ve a c ritic a l th a t p ro life ra tin g c e lls w ith in a tissu e w o u ld h a ve a n
C e ll
6 4
in trin sic a b ility to e n c o u ra g e b lo o d ve sse l g ro w th . B u t c a se a n g io g e n e sis w a s fo u n d to b e a c tiva te d in m id -
sta g e le sio n s, p rio r to th e a p p e a ra n c e o f fu ll-b lo w n tu - th e e vid e n c e is o th e rw ise . T h e c e lls w ith in a b e rra n t p ro -
life ra tive le sio n s in itia lly la c k a n g io g e n ic a b ility, c u rta il- m o rs. S im ila rly, a n g io g e n e sis c a n b e d isc e rn e d in p re -
m a lig n a n t le sio n s o f th e h u m a n c e rvix , b re a st, a n d sk in in g th e ir c a p a b ility fo r e x p a n sio n . I n o rd e r to p ro g re ss
to a la rg e r size , in c ip ie n t n e o p la sia s m u st d e ve lo p a n g io - m e la n o c yte s H a n a h a n a n d F o lk m a n , 1 9 9 6 ; w e e x p e c t
th a t in d u c tio n o f a n g io g e n e sis w ill p ro ve to b e a n e a rly g e n ic a b ility B o u c k e t a l. , 1 9 9 6 ; H a n a h a n a n d F o lk m a n ,
1 9 9 6 ; F o lk m a n , 1 9 9 7 . to m id sta g e e ve n t in m a n y h u m a n c a n c e rs. T h e se o b se r-
va tio n s, ta k e n to g e th e r w ith th e e ffe c ts o f a n g io g e n e sis C o u n te rb a la n c in g p o sitive a n d n e g a tive sig n a ls e n -
c o u ra g e o r b lo c k a n g io g e n e sis. O n e c la ss o f th e se sig - in h ib ito rs, in d ic a te th a t n e o va sc u la riza tio n is a p re re q u i-
site to th e ra p id c lo n a l e x p a n sio n a sso c ia te d w ith th e n a ls is c o n ve ye d b y so lu b le fa c to rs a n d th e ir re c e p to rs,
th e la tte r d isp la ye d o n th e su rfa c e o f e n d o th e lia l c e lls; fo rm a tio n o f m a c ro sc o p ic tu m o rs.
T u m o rs a p p e a r to a c tiva te th e a n g io g e n ic sw itc h b y in te g rin s a n d a d h e sio n m o le c u le s m e d ia tin g c e llm a trix
a n d c e llc e ll a sso c ia tio n a lso p la y c ritic a l ro le s. T h e c h a n g in g th e b a la n c e o f a n g io g e n e sis in d u c e rs a n d
c o u n te rva ilin g in h ib ito rs H a n a h a n a n d F o lk m a n , 1 9 9 6 . a n g io g e n e sis-in itia tin g sig n a ls a re e x e m p lifie d b y va s-
c u la r e n d o th e lia l g ro w th fa c to r VE G F a n d a c id ic a n d O n e c o m m o n stra te g y fo r sh iftin g th e b a la n c e in vo lve s
a lte re d g e n e tra n sc rip tio n . M a n y tu m o rs e vid e n c e in - b a sic fib ro b la st g ro w th fa c to rs F G F 1 /2 . E a c h b in d s to
tra n sm e m b ra n e tyro sin e k in a se re c e p to rs d isp la ye d b y c re a se d e x p re ssio n o f VE G F a n d /o r F G F s c o m p a re d to
th e ir n o rm a l tissu e c o u n te rp a rts. I n o th e rs, e x p re ssio n e n d o th e lia l c e lls F e d i e t a l. , 1 9 9 7 ; Ve ik k o la a n d A lita lo ,
1 9 9 9 . A p ro to typ ic a l a n g io g e n e sis in h ib ito r is th ro m - o f e n d o g e n o u s in h ib ito rs su c h a s th ro m b o sp o n d in -1 o r
-in te rfe ro n is d o w n re g u la te d . M o re o ve r, b o th tra n si- b o sp o n d in -1 , w h ic h b in d s to C D 3 6 , a tra n sm e m b ra n e
re c e p to ro n e n d o th e lia l c e lls c o u p le d to in tra c e llu la rS rc - tio n s m a y o c c u r, a n d in d e e d b e lin k e d , in so m e tu m o rs
S in g h e t a l. , 1 9 9 5 ; Vo lp e rt e t a l. , 1 9 9 7 . lik e tyro sin e k in a se s B u ll e t a l. , 1 9 9 4 . T h e re a re c u r-
re n tly m o re th a n tw o d o ze n a n g io g e n ic in d u c e r fa c to rs T h e m e c h a n ism s u n d e rlyin g sh ifts in th e b a la n c e s b e -
tw e e n a n g io g e n ic re g u la to rs re m a in in c o m p le te ly u n - k n o w n a n d a sim ila r n u m b e r o f e n d o g e n o u s in h ib ito r
p ro te in s. d e rsto o d . I n o n e w e ll-d o c u m e n te d e x a m p le , th e in h ib i-
to r th ro m b o sp o n d in -1 h a s b e e n fo u n d to p o sitive ly I n te g rin sig n a lin g a lso c o n trib u te s to th is re g u la to ry
b a la n c e . Q u ie sc e n t ve sse ls e x p re ss o n e c la ss o f in te - re g u la te d b y th e p 5 3 tu m o r su p p re sso r p ro te in in so m e
c e ll typ e s. C o n se q u e n tly, lo ss o f p 5 3 fu n c tio n , w h ic h g rin s, w h e re a s sp ro u tin g c a p illa rie s e x p re ss a n o th e r.
I n te rfe re n c e w ith sig n a lin g fro m th e la tte r c la ss o f in te - o c c u rs in m o st h u m a n tu m o rs, c a n c a u se th ro m b o sp o n -
d in -1 le ve ls to fa ll, lib e ra tin g e n d o th e lia l c e lls fro m its g rin s c a n in h ib it a n g io g e n e sis Va rn e r a n d C h e re sh ,
1 9 9 6 ; G ia n c o tti a n d R u o sla h ti, 1 9 9 9 , u n d e rsc o rin g th e in h ib ito ry e ffe c ts D a m e ro n e t a l. , 1 9 9 4 . T h e VEGF g e n e
is a lso u n d e r c o m p le x tra n sc rip tio n a l c o n tro l. F o r e x a m - im p o rta n t c o n trib u tio n o f c e ll a d h e sio n to th e a n g io g e n ic
p ro g ra m H yn e s a n d Wa g n e r, 1 9 9 6 . E x tra c e llu la r p ro te - p le , a c tiva tio n o f th e ras o n c o g e n e o r lo ss o f th e VH L
tu m o r su p p re sso r g e n e in c e rta in c e ll typ e s c a u se s a se s a re p h ysic a lly a n d fu n c tio n a lly c o n n e c te d w ith p ro -
a n g io g e n ic in te g rin s, a n d b o th h e lp d ic ta te th e in va sive u p re g u la tio n o f VE G F e x p re ssio n R a k e t a l. , 1 9 9 5 ; M a x -
w e ll e t a l. , 1 9 9 9 . c a p a b ility o f a n g io g e n ic e n d o th e lia l c e lls S te tle r-S te -
ve n so n , 1 9 9 9 . A n o th e r d im e n sio n o f re g u la tio n is e m e rg in g in th e
fo rm o f p ro te a se s, w h ic h c a n c o n tro l th e b io a va ila b ility E x p e rim e n ta l e vid e n c e fo r th e im p o rta n c e o f in d u c in g
a n d su sta in in g a n g io g e n e sis in tu m o rs is b o th e x te n sive o f a n g io g e n ic a c tiva to rs a n d in h ib ito rs. T h u s, a va rie ty
o f p ro te a se s c a n re le a se b F G F sto re d in th e E C M a n d c o m p e llin g B o u c k e t a l. , 1 9 9 6 ; H a n a h a n a n d F o lk -
m a n , 1 9 9 6 ; F o lk m a n , 1 9 9 7 . T h e sto ry b e g in s a lm o st 3 0 Wh ite lo c k e t a l. , 1 9 9 6 , w h e re a s p la sm in , a p ro a n g io -
g e n ic c o m p o n e n t o f th e c lo ttin g syste m , c a n c le a ve itse lf ye a rs a g o w ith F o lk m a n a n d c o lle a g u e s, w h o u se d in
vivo b io a ssa ys to d e m o n stra te th e n e c e ssity o f a n g io - in to a n a n g io g e n e sis in h ib ito r fo rm c a lle d a n g io sta tin
G a te ly e t a l. , 1 9 9 7 . T h e c o o rd in a te d e x p re ssio n o f p ro - g e n e sis fo r e x p lo sive g ro w th o f tu m o r e x p la n ts re -
vie w e d in F o lk m a n , 1 9 9 7 . M o le c u la r p ro o f o f p rin c ip le a n d a n tia n g io g e n ic sig n a lin g m o le c u le s, a n d th e ir m o d -
u la tio n b y p ro te o lysis, a p p e a r to re fle c t th e c o m p le x c a m e , fo r e x a m p le , w h e n a n ti-VE G F a n tib o d ie s p ro ve d
a b le to im p a ir n e o va sc u la riza tio n a n d g ro w th o f su b c u - h o m e o sta tic re g u la tio n o f n o rm a l tissu e a n g io g e n e sis
a n d o f va sc u la r in te g rity. ta n e o u s tu m o rs in m ic e K im e t a l. , 1 9 9 3 , a s d id a d o m i-
n a n t-in te rfe rin g ve rsio n o f th e VE G F re c e p to r 2 flk -1 A s is a lre a d y a p p a re n t, tu m o r a n g io g e n e sis o ffe rs a
u n iq u e ly a ttra c tive th e ra p e u tic ta rg e t, in d e e d o n e th a t M illa u e r e t a l. , 1 9 9 4 ; b o th re su lts h a ve m o tiva te d th e
d e ve lo p m e n t o f sp e c ific VE G F /VE G F -R in h ib ito rs n o w is sh a re d in c o m m o n b y m o st a n d p e rh a p s a ll typ e s o f
h u m a n tu m o rs. T h e n e x t d e c a d e w ill p ro d u c e a c a ta lo g in la te sta g e c lin ic a l tria ls.
T h e e sse n tia l ro le o f a n g io g e n e sis is fu rth e r su p p o rte d o f th e a n g io g e n ic re g u la to ry m o le c u le s e x p re sse d b y
d iffe re n t typ e s o f tu m o rs, a n d in m a n y c a se s, b y th e ir b y th e a b ility o f a n in c re a sin g c a ta lo g o f a n tia n g io g e n ic
su b sta n c e s to im p a ir th e g ro w th o f tu m o r c e lls in o c u - p ro g e n ito r sta g e s. U se o f in c re a sin g ly so p h istic a te d
m o u se m o d e ls w ill m a k e it p o ssib le to a ssig n sp e c ific la te d su b c u ta n e o u sly in m ic e F o lk m a n , 1 9 9 7 . T u m o rs
a risin g in c a n c e r-p ro n e tra n sg e n ic m ic e a re sim ila rly ro le s to e a c h o f th e se re g u la to rs a n d to d isc e rn th e
m o le c u la r m e c h a n ism s th a t g o ve rn th e ir p ro d u c tio n a n d su sc e p tib le to a n g io g e n ic in h ib ito rs B e rg e rs e t a l. ,
1 9 9 9 . a c tivity. A lre a d y a va ila b le e vid e n c e in d ic a te s th a t d iffe r-
e n t typ e s o f tu m o r c e lls u se d istin c t m o le c u la r stra te g ie s T h e a b ility to in d u c e a n d su sta in a n g io g e n e sis se e m s
to b e a c q u ire d in a d isc re te ste p o r ste p s d u rin g tu m o r to a c tiva te th e a n g io g e n ic sw itc h . T h is ra ise s th e q u e s-
tio n o f w h e th e r a sin g le a n tia n g io g e n ic th e ra p e u tic w ill d e ve lo p m e n t, via a n a n g io g e n ic sw itc h fro m va sc u la r
q u ie sc e n c e . Wh e n th re e tra n sg e n ic m o u se m o d e ls w e re su ffic e to tre a t a ll tu m o r typ e s, o r w h e th e r a n e n se m b le
o f su c h th e ra p e u tic s w ill n e e d to b e d e ve lo p e d , e a c h a n a lyze d th ro u g h o u t m u ltiste p tu m o rig e n e sis, in e a c h
R e vie w
6 5
re sp o n d in g to a d istin c t p ro g ra m o f a n g io g e n e sis th a t C h a n g e s in e x p re ssio n o f C A M s in th e im m u n o g lo b u -
lin su p e rfa m ily a lso a p p e a r to p la y c ritic a l ro le s in th e h a s b e e n d e ve lo p e d b y a sp e c ific c la ss o f h u m a n
p ro c e sse s o f in va sio n a n d m e ta sta sis J o h n so n , 1 9 9 1 . tu m o rs.
T h e c le a re st c a se in vo lve s N -C A M , w h ic h u n d e rg o e s a
sw itc h in e x p re ssio n fro m a h ig h ly a d h e sive iso fo rm to
Acquired Capability: Tissue Invasionand Metastasis
p o o rly a d h e sive o r e ve n re p u lsive fo rm s in Wilm s tu -
S o o n e r o r la te r d u rin g th e d e ve lo p m e n t o f m o st typ e s
m o r, n e u ro b la sto m a , a n d sm a ll c e ll lu n g c a n c e r J o h n -
o f h u m a n c a n c e r, p rim a ry tu m o r m a sse s sp a w n p io n e e r
so n , 1 9 9 1 ; K a ise r e t a l. , 1 9 9 6 a n d re d u c tio n in o ve ra ll
c e lls th a t m o ve o u t, in va d e a d ja c e n t tissu e s, a n d th e n c e
e x p re ssio n le ve l in in va sive p a n c re a tic a n d c o lo re c ta l
tra ve l to d ista n t site s w h e re th e y m a y su c c e e d in fo u n d -
c a n c e rs F o g a r e t a l. , 1 9 9 7 . E x p e rim e n ts in tra n sg e n ic
in g n e w c o lo n ie s. T h e se d ista n t se ttle m e n ts o f tu m o r
m ic e su p p o rt a fu n c tio n a l ro le fo r th e n o rm a l a d h e sive
c e lls m e ta sta se s a re th e c a u se o f 9 0 % o f h u m a n c a n -
fo rm o f N -C A M in su p p re ssin g m e ta sta sis P e rl e t a l. ,
c e r d e a th s S p o rn , 1 9 9 6 . T h e c a p a b ility fo r in va sio n a n d
1 9 9 9 .
m e ta sta sis e n a b le s c a n c e r c e lls to e sc a p e th e p rim a ry
C h a n g e s in in te g rin e x p re ssio n a re a lso e vid e n t in
tu m o r m a ss a n d c o lo n ize n e w te rra in in th e b o d y w h e re ,
in va sive a n d m e ta sta tic c e lls. I n va d in g a n d m e ta sta siz-
a t le a st in itia lly, n u trie n ts a n d sp a c e a re n o t lim itin g . T h e
in g c a n c e r c e lls e x p e rie n c e c h a n g in g tissu e m ic ro e n vi-
n e w ly fo rm e d m e ta sta se s a rise a s a m a lg a m s o f c a n c e r
ro n m e n ts d u rin g th e ir jo u rn e ys, w h ic h c a n p re se n t n o ve l
c e lls a n d n o rm a l su p p o rtin g c e lls c o n sc rip te d fro m th e
m a trix c o m p o n e n ts. A c c o rd in g ly, su c c e ssfu l c o lo n iza -
h o st tissu e . L ik e th e fo rm a tio n o f th e p rim a ry tu m o r
tio n o f th e se n e w site s b o th lo c a l a n d d ista n t d e m a n d s
m a ss, su c c e ssfu l in va sio n a n d m e ta sta sis d e p e n d u p o n
a d a p ta tio n , w h ic h is a c h ie ve d th ro u g h sh ifts in th e sp e c -
a ll o f th e o th e r five a c q u ire d h a llm a rk c a p a b ilitie s. B u t
tru m o f in te g rin o r su b u n its d isp la ye d b y th e m ig ra t-
w h a t a d d itio n a l c e llu la r c h a n g e s e n a b le th e a c q u isitio n
in g c e lls. T h e se n o ve l p e rm u ta tio n s re su lt in d iffe re n t
o f th e se fin a l c a p a b ilitie s d u rin g tu m o rig e n e sis?
in te g rin su b typ e s o f w h ic h th e re a re g re a te r th a n 2 2
I n va sio n a n d m e ta sta sis a re e x c e e d in g ly c o m p le x
h a vin g d istin c t su b stra te p re fe re n c e s. T h u s, c a rc in o m a
p ro c e sse s, a n d th e ir g e n e tic a n d b io c h e m ic a l d e te rm i-
c e lls fa c ilita te in va sio n b y sh iftin g th e ir e x p re ssio n o f
n a n ts re m a in in c o m p le te ly u n d e rsto o d . A t th e m e c h a -
in te g rin s fro m th o se th a t fa vo r th e E C M p re se n t in n o r-
n istic le ve l, th e y a re c lo se ly a llie d p ro c e sse s, w h ic h ju sti-
m a l e p ith e liu m to o th e r in te g rin s e . g . , 3 1 a n d V3
fie s th e ir a sso c ia tio n w ith o n e a n o th e r a s o n e g e n e ra l
th a t p re fe re n tia lly b in d th e d e g ra d e d stro m a l c o m p o -
c a p a b ility o f c a n c e r c e lls. B o th u tilize sim ila r o p e ra tio n a l
n e n ts p ro d u c e d b y e x tra c e llu la r p ro te a se s Va rn e r a n d
stra te g ie s, in vo lvin g c h a n g e s in th e p h ysic a l c o u p lin g
C h e re sh , 1 9 9 6 ; L u k a sh e v a n d We rb , 1 9 9 8 . F o rc e d e x -
o f c e lls to th e ir m ic ro e n viro n m e n t a n d a c tiva tio n o f e x -
p re ssio n o f in te g rin su b u n its in c u ltu re d c e lls c a n in d u c e
tra c e llu la r p ro te a se s.
o r in h ib it in va sive a n d m e ta sta tic b e h a vio r, c o n siste n t
S e ve ra l c la sse s o f p ro te in s in vo lve d in th e te th e rin g
w ith a ro le o f th e se re c e p to rs in a c tin g a s c e n tra l d e te r-
o f c e lls to th e ir su rro u n d in g s in a tissu e a re a lte re d in
m in a n ts o f th e se p ro c e sse s Va rn e r a n d C h e re sh , 1 9 9 6 .
c e lls p o sse ssin g in va sive o r m e ta sta tic c a p a b ilitie s. T h e
A tte m p ts a t e x p la in in g th e c e ll b io lo g ic a l e ffe c ts o f
a ffe c te d p ro te in s in c lu d e c e llc e ll a d h e sio n m o le c u le s
in te g rin s in te rm s o f a sm a ll n u m b e r o f m e c h a n istic ru le s
C A M s n o ta b ly m e m b e rs o f th e im m u n o g lo b u lin a n d
h a ve b e e n c o n fo u n d e d b y th e la rg e n u m b e r o f d istin c t
c a lc iu m -d e p e n d e n t c a d h e rin fa m ilie s, b o th o f w h ic h m e -
in te g rin g e n e s, b y th e e ve n la rg e r n u m b e r o f h e te ro d i-
d ia te c e ll-to -c e ll in te ra c tio n s a n d in te g rin s, w h ic h lin k
m e ric re c e p to rs re su ltin g fro m c o m b in a to ria l e x p re ssio n
c e lls to e x tra c e llu la r m a trix su b stra te s. N o ta b ly, a ll o f
o f va rio u s a n d re c e p to r su b u n its, a n d b y th e in c re a s-
th e se a d h e re n c e in te ra c tio n s c o n ve y re g u la to ry sig -
in g e vid e n c e o f c o m p le x sig n a ls e m itte d b y th e c yto -
n a ls to th e c e ll A p lin e t a l. , 1 9 9 8 . T h e m o st w id e ly
p la sm ic d o m a in s o f th e se re c e p to rs A p lin e t a l. , 1 9 9 8 ;
o b se rve d a lte ra tio n in c e ll-to -e n viro n m e n t in te ra c tio n s
G ia n c o tti a n d R u o sla h ti, 1 9 9 9 . S till, th e re is little d o u b t
in c a n c e r in vo lve s E -c a d h e rin , a h o m o typ ic c e ll-to -c e ll
th a t th e se re c e p to rs p la y c e n tra l ro le s in th e c a p a b ility
in te ra c tio n m o le c u le u b iq u ito u sly e x p re sse d o n e p ith e -
fo r tissu e in va sio n a n d m e ta sta sis.
lia l c e lls. C o u p lin g b e tw e e n a d ja c e n t c e lls b y E -c a d h e rin
T h e se c o n d g e n e ra l p a ra m e te r o f th e in va sive a n d
b rid g e s re su lts in th e tra n sm issio n o f a n tig ro w th a n d
m e ta sta tic c a p a b ility in vo lve s e x tra c e llu la r p ro te a se s
o th e r sig n a ls via c yto p la sm ic c o n ta c ts w ith -c a te n in
C o u sse n s a n d We rb , 1 9 9 6 ; C h a m b e rs a n d M a trisia n ,
to in tra c e llu la r sig n a lin g c irc u its th a t in c lu d e th e L e f/
1 9 9 7 . P ro te a se g e n e s a re u p re g u la te d , p ro te a se in h ib i-
T c f tra n sc rip tio n fa c to r C h risto fo ri a n d S e m b , 1 9 9 9 .
to r g e n e s a re d o w n re g u la te d , a n d in a c tive zym o g e n
E -c a d h e rin fu n c tio n is a p p a re n tly lo st in a m a jo rity o f
fo rm s o f p ro te a se s a re c o n ve rte d in to a c tive e n zym e s.
e p ith e lia l c a n c e rs, b y m e c h a n ism s th a t in c lu d e m u ta -
M a trix -d e g ra d in g p ro te a se s a re c h a ra c te ristic a lly a sso -
tio n a l in a c tiva tio n o f th e E -c a d h e rin o r -c a te n in g e n e s,
c ia te d w ith th e c e ll su rfa c e , b y syn th e sis w ith a tra n s-
tra n sc rip tio n a l re p re ssio n , o r p ro te o lysis o f th e e x tra c e l-
m e m b ra n e d o m a in , b in d in g to sp e c ific p ro te a se re -
lu la r c a d h e rin d o m a in C h risto fo ri a n d S e m b , 1 9 9 9 .
c e p to rs, o r a sso c ia tio n w ith in te g rin s We rb , 1 9 9 7 ;
F o rc e d e x p re ssio n o f E -c a d h e rin in c u ltu re d c a n c e r c e lls
S te tle r-S te ve n so n , 1 9 9 9 . O n e im a g in e s th a t d o c k in g o f
a n d in a tra n sg e n ic m o u se m o d e l o f c a rc in o g e n e sis im -
a c tive p ro te a se s o n th e c e ll su rfa c e c a n fa c ilita te in va -
p a irs in va sive a n d m e ta sta tic p h e n o typ e s, w h e re a s in -
sio n b y c a n c e r c e lls in to n e a rb y stro m a , a c ro ss b lo o d
te rfe re n c e w ith E -c a d h e rin fu n c tio n e n h a n c e s b o th ve sse l w a lls, a n d th ro u g h n o rm a l e p ith e lia l c e ll la ye rs.
c a p a b ilitie s C h risto fo ri a n d S e m b , 1 9 9 9 . T h u s, E -c a d - T h a t n o tio n n o tw ith sta n d in g , it is d iffic u lt to u n a m b ig u -
h e rin se rve s a s a w id e ly a c tin g su p p re sso r o f in va sio n o u sly a sc rib e th e fu n c tio n s o f p a rtic u la r p ro te a se s so le ly
a n d m e ta sta sis b y e p ith e lia l c a n c e rs, a n d its fu n c tio n a l to th is c a p a b ility, g ive n th e ir e vid e n t ro le s in o th e r
e lim in a tio n re p re se n ts a k e y ste p in th e a c q u isitio n o f h a llm a rk c a p a b ilitie s, in c lu d in g a n g io g e n e sis S te tle r-
S te ve n so n , 1 9 9 9 a n d g ro w th sig n a lin g We rb , 1 9 9 7 ; th is c a p a b ility.
C e ll
6 6
F ig u re 4 . P a ra lle l P a th w a ys o f T u m o rig e n -
e sis
Wh ile w e b e lie ve th a t virtu a lly a ll c a n c e rs
m u st a c q u ire th e sa m e six h a llm a rk c a p a b ili-
tie s A , th e ir m e a n s o f d o in g so w ill va ry sig -
n ific a n tly, b o th m e c h a n istic a lly se e te x t a n d
c h ro n o lo g ic a lly B . T h u s, th e o rd e r in w h ic h
th e se c a p a b ilitie s a re a c q u ire d se e m s lik e ly
b e q u ite va ria b le a c ro ss th e sp e c tru m o f c a n -
c e r typ e s a n d su b typ e s. M o re o ve r, in so m e
tu m o rs, a p a rtic u la r g e n e tic le sio n m a y c o n fe r
se ve ra l c a p a b ilitie s sim u lta n e o u sly, d e c re a s-
in g th e n u m b e r o f d istin c t m u ta tio n a l ste p s
re q u ire d to c o m p le te tu m o rig e n e sis. T h u s,
lo ss o f fu n c tio n o f th e p 5 3 tu m o r su p p re sso r
c a n fa c ilita te b o th a n g io g e n e sis a n d re sis-
ta n c e to a p o p to sis e . g . , in th e five -ste p p a th -
w a y sh o w n , a s w e ll a s e n a b lin g th e c h a ra c -
te ristic o f g e n o m ic in sta b ility. I n o th e r tu m o rs,
a c a p a b ility m a y o n ly b e a c q u ire d th ro u g h th e
c o lla b o ra tio n o f tw o o r m o re d istin c t g e n e tic
c h a n g e s, th e re b y in c re a sin g th e to ta l n u m b e r
n e c e ssa ry fo r c o m p le tio n o f tu m o r p ro g re s-
sio n . T h u s, in th e e ig h t-ste p p a th w a y sh o w n ,
in va sio n /m e ta sta sis a n d re sista n c e to a p o -
p to sis a re e a c h a c q u ire d in tw o ste p s.
B e rg e rs a n d C o u sse n s, 2 0 0 0 , w h ic h in tu rn c o n trib u te T h e a va ila b le e vid e n c e su g g e sts th a t m o st a re a c q u ire d ,
d ire c tly o r in d ire c tly, th ro u g h c h a n g e s in th e g e n o m e s d ire c tly o r in d ire c tly to th e in va sive /m e ta sta tic c a p a -
b ility. o f c a n c e r c e lls. B u t m u ta tio n o f sp e c ific g e n e s is a n
in e ffic ie n t p ro c e ss, re fle c tin g th e u n c e a sin g , fa stid io u s A fu rth e r d im e n sio n o f c o m p le x ity d e rive s fro m th e
m u ltip le c e ll typ e s in vo lve d in p ro te a se e x p re ssio n a n d m a in te n a n c e o f g e n o m ic in te g rity b y a c o m p le x a rra y
o f D N A m o n ito rin g a n d re p a ir e n zym e s. T h e se g e n o m e d isp la y. I n m a n y typ e s o f c a rc in o m a s, m a trix -d e g ra d in g
p ro te a se s a re p ro d u c e d n o t b y th e e p ith e lia l c a n c e r c e lls m a in te n a n c e te a m s strive to e n su re th a t D N A se q u e n c e
in fo rm a tio n re m a in s p ristin e . K a ryo typ ic o rd e ris g u a ra n - b u t ra th e rb y c o n sc rip te d stro m a l a n d in fla m m a to ry c e lls
We rb , 1 9 9 7 ; o n c e re le a se d b y th e se c e lls, th e y m a y b e te e d b y ye t o th e r w a tc h m e n , m a n n in g so -c a lle d c h e c k -
p o in ts, th a t o p e ra te a t c ritic a l tim e s in th e c e lls life , w ie ld e d b y th e c a rc in o m a c e lls. F o r e x a m p le , c e rta in
c a n c e r c e lls in d u c e u ro k in a se u P A e x p re ssio n in c o c u l- n o ta b ly m ito sis. T o g e th e r, th e se syste m s e n su re th a t
tu re d stro m a l c e lls, w h ic h th e n b in d s to th e u ro k in a se m u ta tio n s a re ra re e ve n ts, in d e e d so ra re th a t th e m u lti-
re c e p to r u P A R e x p re sse d o n th e c a n c e r c e lls J o h n se n p le m u ta tio n s k n o w n to b e p re se n t in tu m o r c e ll g e -
e t a l. , 1 9 9 8 . n o m e s a re h ig h ly u n lik e ly to o c c u r w ith in a h u m a n life
T h e a c tiva tio n o f e x tra c e llu la r p ro te a se s a n d th e a l- sp a n .
te re d b in d in g sp e c ific itie s o f c a d h e rin s, C A M s, a n d in te - Ye t c a n c e rs d o a p p e a r a t su b sta n tia l fre q u e n c y in
g rin s a re c le a rly c e n tra l to th e a c q u isitio n o f in va sive - th e h u m a n p o p u la tio n , c a u sin g so m e to a rg u e th a t th e
n e ss a n d m e ta sta tic a b ility. B u t th e re g u la to ry c irc u its g e n o m e s o f tu m o r c e lls m u st a c q u ire in c re a se d m u ta b il-
a n d m o le c u la r m e c h a n ism s th a t g o ve rn th e se sh ifts re - ity in o rd e r fo r th e p ro c e ss o f tu m o r p ro g re ssio n to re a c h
m a in e lu sive a n d , a t p re se n t, se e m to d iffe r fro m o n e c o m p le tio n in se ve ra l d e c a d e s tim e L o e b , 1 9 9 1 . M a l-
tissu e e n viro n m e n t to a n o th e r. T h e a c q u ire d c a p a b ility fu n c tio n o f sp e c ific c o m p o n e n ts o f th e se g e n o m ic
fo r in va sio n a n d m e ta sta sis re p re se n ts th e la st g re a t c a re ta k e r syste m s h a s b e e n in vo k e d to e x p la in th is
fro n tie r fo r e x p lo ra to ry c a n c e r re se a rc h . We e n visio n in c re a se d m u ta b ility L e n g a u e r e t a l. , 1 9 9 8 . T h e m o st
th a t e vo lvin g a n a lytic te c h n iq u e s w ill so o n m a k e it p o ssi- p ro m in e n t m e m b e r o f th e se syste m s is th e p 5 3 tu m o r
b le to c o n stru c t c o m p re h e n sive p ro file s o f th e e x p re s- su p p re sso r p ro te in , w h ic h , in re sp o n se to D N A d a m a g e ,
sio n a n d fu n c tio n a l a c tivitie s o f p ro te a se s, in te g rin s, a n d e lic its e ith e r c e ll c yc le a rre st to a llo w D N A re p a ir to ta k e
C A M s in a w id e va rie ty o f c a n c e r typ e s, b o th b e fo re a n d p la c e o r a p o p to sis if th e d a m a g e is e x c e ssive . I n d e e d , it
a fte r th e y a c q u ire in va sive a n d m e ta sta tic a b ilitie s. T h e is n o w c le a r th a t th e fu n c tio n in g o f th e p 5 3 D N A d a m a g e
c h a lle n g e w ill th e n b e to a p p ly th e n e w m o le c u la r in - sig n a lin g p a th w a y is lo st in m o st, if n o t a ll, h u m a n c a n -
sig h ts a b o u t tissu e in va sive n e ss a n d m e ta sta sis to th e c e rs L e vin e , 1 9 9 7 . M o re o ve r, a g ro w in g n u m b e r o f
d e ve lo p m e n t o f e ffe c tive th e ra p e u tic stra te g ie s. o th e r g e n e s in vo lve d in se n sin g a n d re p a irin g D N A d a m -
a g e , o r in a ssu rin g c o rre c t c h ro m o so m a l se g re g a tio n
d u rin g m ito sis, is fo u n d to b e lo st in d iffe re n t c a n c e rs, AnEnabling Characteristic: Genome Instability
T h e a c q u isitio n o f th e e n u m e ra te d six c a p a b ilitie s d u rin g la b e lin g th e se c a re ta k e rs a s tu m o r su p p re sso rs L e n -
g a u e r e t a l. , 1 9 9 8 . T h e ir lo ss o f fu n c tio n is e n visio n e d th e c o u rse o f tu m o r p ro g re ssio n c re a te s a d ile m m a .
R e vie w
6 7
to a llo w g e n o m e in sta b ility a n d va ria b ility a n d th e g e n e r- th e sig n a ls e x c h a n g e d b e tw e e n th e va rio u s c e ll typ e s
e x istin g sym b io tic a lly w ith in a tu m o r m a ss a n d k n o w in g a tio n o f c o n se q u e n tly m u ta n t c e lls w ith se le c tive a d va n -
ta g e s. I n te re stin g ly, re c e n t e vid e n c e su g g e sts th a t a p o - th e ir e ffe c ts o n th e in te g ra te d c irc u its o f e a c h o f th o se
c e ll typ e s. p to sis m a y a lso b e a ve h ic le o f g e n o m ic in sta b ility, in
th a t D N A w ith in a p o p to tic c e ll b o d ie s c a n b e in c o rp o - O u r a b ility to a n a lyze in d ivid u a l h u m a n c a n c e rs a t
th e g e n e tic a n d b io c h e m ic a l le ve ls w ill a lso u n d e rg o a ra te d in to n e ig h b o rin g c e lls fo llo w in g p h a g o c tyto sis
H o lm g re n e t a l. , 1 9 9 9 , in p rin c ip le g e n e tic a lly d ive rsify- d ra m a tic c h a n g e . A t p re se n t, d e sc rip tio n o f a re c e n tly
d ia g n o se d tu m o r in te rm s o f its u n d e rlyin g g e n e tic le - in g a n y o f th e c o n stitu e n t c e ll typ e s o f a tu m o r. We p la c e
th is a c q u ire d c h a ra c te ristic o f g e n o m ic in sta b ility a p a rt sio n s re m a in s a d ista n t p ro sp e c t. N o n e th e le ss, w e lo o k
a h e a d 1 0 o r 2 0 ye a rs to th e tim e w h e n th e d ia g n o sis o f fro m th e six a c q u ire d c a p a b ilitie s a sso c ia te d w ith tu m o r
c e ll p h e n o typ e a n d tu m o r p h ysio lo g y: it re p re se n ts th e a ll th e so m a tic a lly a c q u ire d le sio n s p re se n t in a tu m o r
c e ll g e n o m e w ill b e c o m e a ro u tin e p ro c e d u re . B y th e n , m e a n s th a t e n a b le s e vo lvin g p o p u la tio n s o f p re m a lig -
n a n t c e lls to re a c h th e se six b io lo g ic a l e n d p o in ts. g e n o m e -w id e g e n e e x p re ssio n p ro file s o f tu m o r c e lls
w ill a lso b e ro u tin e . With a ll th is in fo rm a tio n in h a n d , it
w ill b e c o m e p o ssib le to te st d e fin itive ly o u r p ro p o sitio n Alternative Pathways to Cancer
th a t th e d e ve lo p m e n t o f a ll typ e s o f h u m a n tu m o r c e lls T h e p a th s th a t c e lls ta k e o n th e ir w a y to b e c o m in g m a lig -
is g o ve rn e d b y a c o m m o n se t o f ru le s su c h a s th o se n a n t a re h ig h ly va ria b le . With in a g ive n c a n c e r typ e ,
im p lie d b y th e six a c q u ire d c a p a b ilitie s e n u m e ra te d m u ta tio n o f p a rtic u la r ta rg e t g e n e s su c h a s ras o r p53
h e re . m a y b e fo u n d in o n ly a su b se t o f o th e rw ise h isto lo g ic a lly
We a n tic ip a te fa r d e e p e r in sig h t in to th e ro le s p la ye d id e n tic a l tu m o rs. F u rth e r, m u ta tio n s in c e rta in o n c o -
b y in h e rite d a lle le s in c a n c e r su sc e p tib ility a n d p a th o - g e n e s a n d tu m o r su p p re sso r g e n e s c a n o c c u r e a rly in
g e n e sis. A t p re se n t, o u r u n d e rsta n d in g o f th e in te rp la y so m e tu m o r p ro g re ssio n p a th w a ys a n d la te in o th e rs. A s
a t th e c e llu la r le ve l b e tw e e n in h e rite d c a n c e r m o d ifie r a c o n se q u e n c e , th e a c q u isitio n o f b io lo g ic a l c a p a b ilitie s
g e n e s w ith o n c o g e n e s a n d tu m o r su p p re sso r g e n e s th a t su c h a s re sista n c e to a p o p to sis, su sta in e d a n g io g e n -
a re a lte re d so m a tic a lly is ru d im e n ta ry; m o d ifie rs c a n in e sis, a n d u n lim ite d re p lic a tive p o te n tia l c a n a p p e a r a t
p rin c ip le a c t in a n y o f th e c o n stitu e n t c e ll typ e s o f a d iffe re n t tim e s d u rin g th e se va rio u s p ro g re ssio n s. A c -
tu m o r, o r e lse w h e re in th e b o d y, w h e re a s th e c la ssic a l c o rd in g ly, th e p a rtic u la r se q u e n c e in w h ic h c a p a b ilitie s
c a n c e r g e n e s la rg e ly a c t in th e c a n c e r c e lls th e m se lve s. a re a c q u ire d c a n va ry w id e ly, b o th a m o n g tu m o rs o f th e
T h e se g a p s w ill b e b rid g e d in p a rt b y n e w in fo rm a tic s sa m e typ e a n d c e rta in ly b e tw e e n tu m o rs o f d iffe re n t
te c h n o lo g ie s, e n a b lin g u s to p ro c e ss a n d in te rp re t th e typ e s F ig u re 4 . F u rth e rm o re , in c e rta in tu m o rs, a sp e -
in u n d a tio n o f g e n e tic in fo rm a tio n th a t w ill so o n flo w fro m c ific g e n e tic e ve n t m a y, o n its o w n , c o n trib u te o n ly p a r-
a u to m a te d se q u e n c in g in stru m e n ts. N e w te c h n o lo g ie s tia lly to th e a c q u isitio n o f a sin g le c a p a b ility, w h ile in
w ill a lso a id u s in ra tio n a lizin g th e c o m p le x c o n ste lla - o th e rs, th is e ve n t m a y a id in th e sim u lta n e o u s a c q u isi-
tio n s o f in te ra c tin g a lle le s in te rm s o f a syste m a tic s o f tio n o f se ve ra l d istin c t c a p a b ilitie s. N o n e th e le ss, w e b e -
c a n c e r fo rm a tio n o f th e typ e th a t w e p ro p o se h e re . lie ve th a t in d e p e n d e n t o f h o w th e ste p s in th e se g e n e tic
T h e m e ta p h o rs u se d to c o n c e p tu a lize c a n c e r c e ll p a th w a ys a re a rra n g e d , th e b io lo g ic a l e n d p o in ts th a t
fu n c tio n w ill a lso sh ift d ra m a tic a lly. F o r d e c a d e s n o w , a re u ltim a te ly re a c h e d th e h a llm a rk c a p a b ilitie s o f c a n -
w e h a ve b e e n a b le to p re d ic t w ith p re c isio n th e b e h a vio r c e r w ill p ro ve to b e sh a re d in c o m m o n b y a ll typ e s o f
o f a n e le c tro n ic in te g ra te d c irc u it in te rm s o f its c o n stit- tu m o rs.
u e n t p a rts its in te rc o n n e c tin g c o m p o n e n ts, e a c h re -
sp o n sib le fo ra c q u irin g , p ro c e ssin g , a n d e m ittin g sig n a ls
Synthesis
a c c o rd in g to a p re c ise ly d e fin e d se t o f ru le s. T w o d e -
C a n c e r c e lls p ro p a g a te d in c u ltu re a n d d isse c te d in to
c a d e s fro m n o w , h a vin g fu lly c h a rte d th e w irin g d ia -
th e ir m o le c u la r c o m p o n e n ts h a ve yie ld e d m u c h o f th e
g ra m s o f e ve ry c e llu la r sig n a lin g p a th w a y, it w ill b e p o s-
w e a lth o f in fo rm a tio n th a t w e c u rre n tly p o sse ss a b o u t
sib le to la y o u t th e c o m p le te in te g ra te d c irc u it o f th e
th e m o le c u la r p ro c e sse s u n d e rlyin g c a n c e r d e ve lo p -
c e ll u p o n its c u rre n t o u tlin e F ig u re 2 . We w ill th e n b e
m e n t. Ye t b y sim p lifyin g th e n a tu re o f c a n c e r p o r-
a b le to a p p ly th e to o ls o f m a th e m a tic a l m o d e lin g to
tra yin g it a s a c e ll-a u to n o m o u s p ro c e ss in trin sic to th e
e x p la in h o w sp e c ific g e n e tic le sio n s se rve to re p ro g ra m
c a n c e r c e ll th e se e x p e rim e n ta l m o d e ls h a ve tu rn e d
th is in te g ra te d c irc u it in e a c h o f th e c o n stitu e n t c e ll
th e ir b a c k o n a c e n tra l b io lo g ic a l re a lity o f tu m o r fo rm a -
typ e s so a s to m a n ife st c a n c e r.
tio n in vivo : c a n c e rd e ve lo p m e n t d e p e n d s u p o n c h a n g e s
With h o listic c la rity o f m e c h a n ism , c a n c e r p ro g n o sis
in th e h e te ro typ ic in te ra c tio n s b e tw e e n in c ip ie n t tu m o r
a n d tre a tm e n t w ill b e c o m e a ra tio n a l sc ie n c e , u n re c o g -
c e lls a n d th e ir n o rm a l n e ig h b o rs. M o re o ve r, o n c e fo rm e d ,
n iza b le b y c u rre n t p ra c titio n e rs. I t w ill b e p o ssib le to
virtu a lly a ll typ e s o f h u m a n tu m o rs, in c lu d in g th e ir m e ta -
u n d e rsta n d w ith p re c isio n h o w a n d w h y tre a tm e n t re g i- sta tic o u tg ro w th s, c o n tin u e to h a rb o r c o m p le x m ix tu re s
m e n s a n d sp e c ific a n titu m o r d ru g s su c c e e d o r fa il. We o f se ve ra l c e ll typ e s th a t c o lla b o ra te to c re a te m a lig n a n t
e n visio n a n tic a n c e r d ru g s ta rg e te d to e a c h o f th e h a ll- g ro w th F ig u re 3 . T h is re c o n c e p tu a liza tio n o f c a n c e r
m a rk c a p a b ilitie s o f c a n c e r; so m e , u se d in a p p ro p ria te c e ll b io lo g y h a s b e g u n to d rive p ro fo u n d c h a n g e s in
c o m b in a tio n s a n d in c o n c e rt w ith so p h istic a te d te c h n o l- h o w w e stu d y th is d ise a se e x p e rim e n ta lly. C o n tin u in g
o g ie s to d e te c t a n d id e n tify a ll sta g e s o f d ise a se p ro - e lu c id a tio n o f c a n c e r p a th o g e n e sis w ill d e p e n d in c re a s-
g re ssio n , w ill b e a b le to p re ve n t in c ip ie n t c a n c e rs fro m in g ly u p o n h e te ro typ ic o rg a n c u ltu re syste m s in vitro
d e ve lo p in g , w h ile o th e rs w ill c u re p re e x istin g c a n c e rs, a n d e ve rm o re re fin e d m o u se m o d e ls in vivo . L o o k in g
e lu sive g o a ls a t p re se n t. O n e d a y, w e im a g in e th a t c a n - a h e a d in to th e fu tu re , th e se syste m s w ill h e lp u s c h a rt
c e r b io lo g y a n d tre a tm e n t a t p re se n t, a p a tc h w o rk q u ilt c o m p re h e n sive m a p s o f g ro w th sig n a lin g n e tw o rk s in
c a n c e r, a n e n d e a vo r th a t w ill d e p e n d o n d e fin in g a ll o f o f c e ll b io lo g y, g e n e tic s, h isto p a th o lo g y, b io c h e m istry,
C e ll
6 8
m o le c u le E -c a d h e rin a s a tu m o u r-su p p re sso r g e n e . T re n d s B io -
im m u n o lo g y, a n d p h a rm a c o lo g y w ill b e c o m e a sc i-
c h e m . S c i. 24, 7 3 7 6 .
e n c e w ith a c o n c e p tu a l stru c tu re a n d lo g ic a l c o h e re n c e
C h risto fo ri, G . , N a ik , P . , a n d H a n a h a n , D . 1 9 9 4 . A se c o n d sig n a l
th a t riva ls th a t o f c h e m istry o r p h ysic s.
su p p lie d b y in su lin -lik e g ro w th fa c to r I I in o n c o g e n e -in d u c e d tu m o ri-
g e n e sis. N a tu re 369, 4 1 4 4 1 8 .
Acknowledgments
C o rd o n -C a rd o , C . , a n d P rive s, C . 1 9 9 9 . A t th e c ro ssro a d s o f in fla m -
m a tio n a n d tu m o rig e n e sis. J . E x p . M e d . 190, 1 3 6 7 1 3 7 0 .
We w ish to th a n k T e rry S c h o o p o f B io m e d A rts A sso c ia te s, S a n
C o u n te r, C . M . , A vilio n , A . A . , L e F e u vre , C . E . , S te w a rt, N . G . , G re id e r,
F ra n c isc o , fo r p re p a ra tio n o f th e fig u re s, C o ri B a rg m a n n a n d Z e n a
C . W. , H a rle y, C . B . , a n d B a c c h e tti, S . 1 9 9 2 . T e lo m e re sh o rte n in g
We rb fo r in sig h tfu l c o m m e n ts o n th e m a n u sc rip t, a n d N o rm ita S a n -
a sso c ia te d w ith c h ro m o so m e in sta b ility is a rre ste d in im m o rta l c e lls
to re fo r e d ito ria l a ssista n c e . I n a d d itio n , w e a re in d e b te d to J o e
w h ic h e x p re ss te lo m e ra se a c tivity. E M B O J . 11, 1 9 2 1 1 9 2 9 .
H a rfo rd a n d R ic h a rd K la u sn e r, w h o a llo w e d u s to a d a p t a n d e x p a n d
C o u n te r, C . M . , H a h n , W. C . , We i, W. , D ic k in so n C a d d le , S . , B e ije rs-
th e ir d e p ic tio n o f th e c e ll sig n a lin g n e tw o rk , a n d w e a p p re c ia te
b e rg e n , R . L . , L a n sd o rp , P . M . , S e d ivy, J . M . , a n d We in b e rg , R . A .
su g g e stio n s o n sig n a lin g p a th w a ys fro m R a n d y Wa tn ic k , B ria n E le n -
1 9 9 8 . D isso c ia tio n b e tw e e n te lo m e ra se a c tivity, te lo m e re m a in te -
b a s, B ill L u n d b e rg , D a ve M o rg a n , a n d H e n ry B o u rn e . R . A . W. is
n a n c e a n d c e llu la r im m o rta liza tio n . P ro c . N a tl. A c a d . S c i. U S A 95,
a L u d w ig F o u n d a tio n a n d A m e ric a n C a n c e r S o c ie ty P ro fe sso r o f
1 4 7 2 3 1 4 7 2 8 .
B io lo g y. H is w o rk h a s b e e n su p p o rte d b y th e D e p a rtm e n t o f th e
C o u sse n s, L . M . , a n d We rb , Z . 1 9 9 6 . M a trix m e ta llo p ro te in a se s a n d
A rm y a n d th e N a tio n a l I n stitu te s o f H e a lth . D . H . a c k n o w le d g e s
th e d e ve lo p m e n t o f c a n c e r. C h e m . B io l. 3, 8 9 5 9 0 4 .
th e su p p o rt a n d e n c o u ra g e m e n t o f th e N a tio n a l C a n c e r I n stitu te .
C o u sse n s, L . M . , R a ym o n d , W. W. , B e rg e rs, G . , L a ig -We b ste r, M . ,
E d ito ria l p o lic y h a s re n d e re d th e c ita tio n s illu stra tive b u t n o t c o m -
B e h re n d tse n , O . , We rb , Z . , C a u g h e y, G . H . , a n d H a n a h a n , D . 1 9 9 9 .
p re h e n sive .
I n fla m m a to ry m a st c e lls u p -re g u la te a n g io g e n e sis d u rin g sq u a m o u s
e p ith e lia l c a rc in o g e n e sis. G e n e s D e v. 13, 1 3 8 2 1 3 9 7 .
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R e vie w
6 9
H a h n , W. C . , C o u n te r, C . M . , L u n d b e rg , A . S . , B e ije rsb g e rn , R . L . , M a rk o w itz, S . , Wa n g , J . , M e ye ro ff, L . , P a rso n s, R . , S u n , L . , L u tte r-
b a u g h , J . , F a n , R . , Z b o ro w sk a , E . , K in zle r, K . , Vo g e lste in , B . , e t a l. B ro o k s, M . W. , a n d We in b e rg , R . A . 1 9 9 9 . C re a tio n o f h u m a n tu m o r
c e lls w ith d e fin e d g e n e tic e le m e n ts. N a tu re 400, 4 6 4 4 6 8 . 1 9 9 5 . I n a c tiva tio n o f th e typ e I I T G F - re c e p to r in c o lo n c a n c e r
c e lls w ith m ic ro sa te llite in sta b ility. S c ie n c e 268, 1 3 3 6 1 3 3 8 .
H a lvo rse n , T . L . , L e ib o w itz, G . , a n d L e vin e , F . 1 9 9 9 . T e lo m e ra se
a c tivity is su ffic ie n t to a llo w tra n sfo rm e d c e lls to e sc a p e fro m c risis. M a x w e ll, P . H . , Wie se n e r, M . S . , C h a n g , G . -W. , C liffo rd , S . C . , Va u x ,
E . C . , C o c k m a n , M . E . , Wyk o ff, C . C . , P u g h , C . W. , M a h e r, E . R . , a n d M o l. C e ll. B io l. 19, 1 8 6 4 1 8 7 0 .
R a tc liffe , P . J . 1 9 9 9 . T h e tu m o u r su p p re sso r p ro te in VH L ta rg e ts
H a n a h a n , D . , a n d F o lk m a n , J . 1 9 9 6 . P a tte rn s a n d e m e rg in g m e c h a -
h yp o x ia -in d u c ib le fa c to rs fo r o x yg e n -d e p e n d e n t p ro te o lysis. N a tu re
n ism s o f th e a n g io g e n ic sw itc h d u rin g tu m o rig e n e sis. C e ll 86,
399, 2 7 1 2 7 5 .
3 5 3 3 6 4 .
M c D o n n e ll, T . J . , a n d K o rsm e ye r, S . J . 1 9 9 1 . P ro g re ssio n fro m
H a n n o n , G . J . , a n d B e a c h , D . 1 9 9 4 . P 1 5
I N K 4 B
is a p o te n tia l e ffe c to r
lym p h o id h yp e rp la sia to h ig h -g ra d e m a lig n a n t lym p h o m a in m ic e
o f T G F -b e ta -in d u c e d c e ll c yc le a rre st. N a tu re 371, 2 5 7 2 6 1 .
tra n sg e n ic fo r th e t 1 4 ; 1 8 . N a tu re 349, 2 5 4 2 5 6 .
H a rris, C . C . 1 9 9 6 . p 5 3 tu m o r su p p re sso r g e n e : fro m th e b a sic
M e d e m a , R . H . , a n d B o s, J . L . 1 9 9 3 . T h e ro le o f p 2 1 -ra s in re c e p to r
re se a rc h la b o ra to ry to th e c lin ic a n a b rid g e d h isto ric a l p e rsp e c -
tyro sin e k in a se sig n a lin g . C rit. R e v. O n c o g . 4, 6 1 5 6 6 1 .
tive . C a rc in o g e n e sis 17, 1 1 8 7 1 1 9 8 .
M illa u e r, B . , S h a w ve r, L . K . , P la te , K . H . , R isa u , W. , a n d U llric h , A .
H a yflic k , L . 1 9 9 7 . M o rta lity a n d im m o rta lity a t th e c e llu la r le ve l. A
1 9 9 4 . G lio b la sto m a g ro w th in h ib ite d in vivo b y a d o m in a n t-n e g a tive
re vie w . B io c h e m istry 62, 1 1 8 0 1 1 9 0 .
F lk -1 m u ta n t. N a tu re 367, 5 7 6 5 7 9 .
H o lm g re n , L . , S ze le s, A . , R a jn a vo lg yi, E . , F o lk m a n , J . , K le in , G . ,
M o se s, H . L . , Ya n g , E . Y. , a n d P ie te n p o l, J . A . 1 9 9 0 . T G F - stim u la -
E rn b e rg , I . , a n d F a lk , K . I . 1 9 9 9 . H o rizo n ta l tra n sfe r o f D N A b y th e
tio n a n d in h ib itio n o f c e ll p ro life ra tio n : n e w m e c h a n istic in sig h ts.
u p ta k e o f a p o p to tic b o d ie s. B lo o d 93, 3 9 5 6 3 9 6 3 .
C e ll 63, 2 4 5 2 4 7 .
H u d so n , J . D . , S h o a ib i, M . A . , M a e stro , R . , C a rn e ro , A . , H a n n o n , G . J . ,
N o w e ll, P . C . 1 9 7 6 . T h e c lo n a l e vo lu tio n o f tu m o r c e ll p o p u la tio n s.
a n d B e a c h , D . H . 1 9 9 9 . A p ro in fla m m a to ry c yto k in e in h ib its p 5 3
S c ie n c e 194, 2 3 2 8 .
tu m o r su p p re sso r a c tivity. J . E x p . M e d . 190, 1 3 7 5 1 3 8 2 .
O lu m i, A . F . , G ro ssfe ld , G . D . , H a yw a rd , S . W. , C a rro ll, P . R . , T lsty, T . D . ,
H u e b e r, A . O . , Z o rn ig , M . , L yo n , D . , S u d a , T . , N a g a ta , S . , a n d E va n ,
a n d C u n h a , G . R . 1 9 9 9 . C a rc in o m a -a sso c ia te d fib ro b la sts d ire c t
G . I . 1 9 9 7 . R e q u ire m e n t fo r th e C D 9 5 re c e p to r-lig a n d p a th w a y in
tu m o r p ro g re ssio n o f in itia te d h u m a n p ro sta tic e p ith e liu m . C a n c e r
c -M yc -in d u c e d a p o p to sis. S c ie n c e 278, 1 3 0 5 1 3 0 9 .
R e s. 59, 5 0 0 2 5 0 1 1 .
H u n te r, T . 1 9 9 7 . O n c o p ro te in n e tw o rk s. C e ll 88, 3 3 3 3 4 6 .
P e rl, A . -K . , D a h l, U . , Wilg e n b u s, P . , C re m e r, H . , S e m b , H . , a n d
H yn e s, R . O . , a n d Wa g n e r, D . D . 1 9 9 6 . G e n e tic m a n ip u la tio n o f va s-
C h risto fo ri, G . 1 9 9 9 . R e d u c e d e x p re sio n o f n e u ra l c e ll a d h e sio n
c u la r a d h e sio n m o le c u le s in m ic e . J . C lin . I n ve st. 98, 2 1 9 3 2 1 9 5 .
m o le c u le in d u c e s m e ta sta tic d isse m in a tio n o f p a n c re a tic tu m o r
I sh iza k i, Y. , C h e n g , L . , M u d g e , A . W. , a n d R a ff, M . C . 1 9 9 5 . P ro -
c e lls. N a t. M e d . 5, 2 8 6 2 9 1 .
g ra m m e d c e ll d e a th b y d e fa u lt in e m b ryo n ic c e lls, fib ro b la sts, a n d
P itti, R . M . , M a rste rs, S . A . , L a w re n c e , D . A . , R o y, M . , K isc h k e l, F . C . ,
c a n c e r c e lls. M o l. B io l. C e ll 6, 1 4 4 3 1 4 5 8 .
D o w d , P . , H u a n g , A . , D o n a h u e , C . J . , S h e rw o o d , S . W. , B a ld w in , D . T . ,
J o h n se n , M . , L u n d , L . R . , R o m e r, J . , A lm h o lt, K . , a n d D a n o K . 1 9 9 8 .
e t a l. 1 9 9 8 . G e n o m ic a m p lific a tio n o f a d e c o y re c e p to r fo r F a s
C a n c e r in va sio n a n d tissu e re m o d e lin g : c o m m o n th e m e s in p ro te o -
lig a n d in lu n g a n d c o lo n c a n c e r. N a tu re 396, 6 9 9 7 0 3 .
lytic m a trix d e g ra d a tio n . C u rr. O p in . C e ll B io l. 10, 6 6 7 6 7 1 .
R a k , J . , F ilm u s, J . , F in k e n ze lle r, G . , G ru g e l, S . , M a rm e , D . , a n d K e r-
J o h n so n , J . P . 1 9 9 1 . C e ll a d h e sio n m o le c u le s o f th e im m u n o g lo b u -
b e l, R . S . 1 9 9 5 . O n c o g e n e s a s in d u c e rs o f tu m o r a n g io g e n e sis.
lin su p e rg e n e fa m ily a n d th e ir ro le in m a lig n a n t tra n sfo rm a tio n a n d
C a n c e r M e ta sta sis R e v. 14, 2 6 3 2 7 7 .
p ro g re ssio n to m e ta sta tic d ise a se . C a n c e r M e ta sta sis R e v. 10,
R e n a n , M . J . 1 9 9 3 . H o w m a n y m u ta tio n s a re re q u ire d fo r tu m o rig e n -
1 1 2 2 .
e sis? I m p lic a tio n s fro m h u m a n c a n c e r d a ta . M o l. C a rc in o g e n e sis 7,
K a h n , P . , F ryk b e rg , L . , B ra d y, C . , S ta n le y, I . , B e u g , H . , Ve n n strom ,
1 3 9 1 4 6 .
B . , a n d G ra f, T . 1 9 8 6 . v-erbA c o o p e ra te s w ith sa rc o m a o n c o g e n e s
R o m m e l, C . , a n d H a fe n , E . 1 9 9 8 . R a s a ve rsa tile c e llu la r sw itc h .
in le u k e m ic c e ll tra n sfo rm a tio n . C e ll 45, 3 4 9 3 5 6 .
C u rr. O p in . G e n e t. D e v. 8, 4 1 2 4 1 8 .
K a ise r, U . , A u e rb a c h , B . , a n d O ld e n b u rg , M . 1 9 9 6 . T h e n e u ra l c e ll
S c h u tte , M . , H ru b a n , R . , H e d ric k , L . , C h o , K . , N a d a sd y, G . , We in ste in ,
a d h e sio n m o le c u le N C A M in m u ltip le m ye lo m a . L e u k . L ym p h o m a
C . , B o va , G . , I sa a c s, W. , C a irn s, P . , N a w ro z, H . , e t a l. 1 9 9 6 . DPC4
20, 3 8 9 3 9 5 .
g e n e in va rio u s tu m o r typ e s. C a n c e r R e s. 56, 2 5 2 7 2 5 3 0 .
K e rr, J . F . , Wyllie , A . H . , a n d C u rrie , A . R . 1 9 7 2 . A p o p to sis: a b a sic
S e rra n o , M . , L in , A . W. , M c C u rra c h , M . E . , B e a c h , D . , a n d L o w e , S . W.
b io lo g ic a l p h e n o m e n o n w ith w id e -ra n g in g im p lic a tio n s in tissu e k i-
1 9 9 7 . O n c o g e n e ic ras p ro vo k e s p re m a tu re c e ll se n e sc e n c e a sso c i-
n e tic s. B r. J . C a n c e r 26, 2 3 9 2 5 7 .
a te d w ith a c c u m u la tio n o f p 5 3 a n d p 1 6
I N K 4 A
. C e ll 88, 5 9 3 6 0 2 .
K im , K . J . , L i, B . , Win e r, J . , A rm a n in i, M . , G ille tt, N . , P h ilip p s, H . S . , a n d
S h ib a ta , M . A . , M a ro u la k o u , I . G . , J o rc yk , C . L . , G o ld , L . G . , Wa rd , J . M . ,
F e rra ra , N . 1 9 9 3 . I n h ib itio n o f va sc u la r e n d o th e lia l g ro w th fa c to r-
a n d G re e n , J . E . 1 9 9 6 . p 5 3 -in d e p e n d e n t a p o p to sis d u rin g m a m m a ry
in d u c e d a n g io g e n e sis su p p re sse s tu m o u r g ro w th in vivo . N a tu re
tu m o r p ro g re ssio n in C 3 1 /S V4 0 la rg e T a n tig e n tra n sg e n ic m ic e :
362, 8 4 1 8 4 4 .
su p p re ssio n o f a p o p to sis d u rin g th e tra n sitio n fro m p re n e o p la sia
K in zle r, K . W. , a n d Vo g e lste in , B . 1 9 9 6 . L e sso n s fro m h e re d ita ry
to c a rc in o m a . C a n c e r R e s. 56, 2 9 9 8 3 0 0 3 .
c o lo re c ta l c a n c e r. C e ll 87, 1 5 9 1 7 0 .
S h a y, J . W. , a n d B a c c h e tti, S . 1 9 9 7 . A su rve y o f te lo m e ra se a c tivity
K in zle r, K . W. , a n d Vo g e lste in , B . 1 9 9 8 . L a n d sc a p in g th e c a n c e r
in h u m a n c a n c e r. E u r. J . C a n c e r 33, 7 8 7 7 9 1 .
te rra in . S c ie n c e 280, 1 0 3 6 1 0 3 7 .
S in g h , R . K . , G u tm a n , M . , B u c a n a , C . D . , S a n c h e z, R . , L la n sa , N . ,
K o rsm e ye r, S . J . 1 9 9 2 . C h ro m o so m a l tra n slo c a tio n s in lym p h o id
a n d F id le r, I . J . 1 9 9 5 . I n te rfe ro n s a lp h a a n d b e ta d o w n -re g u la te th e
m a lig n a n c ie s re ve a l n o ve l p ro to -o n c o g e n e s. A n n u . R e v. I m m u n o l.
e x p re ssio n o f b a sic fib ro b la st g ro w th fa c to r in h u m a n c a rc in o m a s.
10, 7 8 5 8 0 7 .
P ro c . N a tl. A c a d . S c i. U S A 92, 4 5 6 2 4 5 6 6 .
L e n g a u e r, C . , K in zle r, K . W. , a n d Vo g e lste in , B . 1 9 9 8 . G e n e tic in sta -
S k o b e , M . , a n d F u se n ig , N . E . 1 9 9 8 . T u m o rig e n ic c o n ve rsio n o f
b ilitie s in h u m a n c a n c e rs. N a tu re 396, 6 4 3 6 4 9 .
im m o rta l h u m a n k e ra tin o c yte s th ro u g h stro m a l c e ll a c tiva tio n . P ro c .
N a tl. A c a d . S c i. U S A 95, 1 0 5 0 1 0 5 5 . L e vin e , A . J . 1 9 9 7 . p 5 3 , th e c e llu la r g a te k e e p e r fo r g ro w th a n d d ivi-
sio n . C e ll 88, 3 2 3 3 3 1 .
S la m o n , D . J . , C la rk , G . M . , Wo n g , S . G . , L e vin , W. J . , U llric h , A . , a n d
M c G u ire , W. L . 1 9 8 7 . H u m a n b re a st c a n c e r: c o rre la tio n o f re la p se L o e b , L . A . 1 9 9 1 . M u ta to r p h e n o typ e m a y b e re q u ire d fo r m u ltiste p
c a rc in o g e n e sis. C a n c e r R e s. 51, 3 0 7 3 3 0 7 9 . a n d su rviva l w ith a m p lific a tio n o f th e H E R -2 /neuo n c o g e n e . S c ie n c e
235, 1 7 7 1 8 2 .
L o te m , J . , a n d S a c h s, L . 1 9 9 6 . C o n tro l o f a p o p to sis in h e m a to p o ie -
sis a n d le u k e m ia b y c yto k in e s, tu m o r su p p re sso r a n d o n c o g e n e s. S p o rn , M . B . 1 9 9 6 . T h e w a r o n c a n c e r. L a n c e t 347, 1 3 7 7 1 3 8 1 .
L e u k e m ia 10, 9 2 5 9 3 1 .
S te tle r-S te ve n so n , W. G . 1 9 9 9 . M a trix m e ta llo p ro te in a se s in a n g io -
g e n e sis: a m o vin g ta rg e t fo r th e ra p e u tic in te rve n tio n . J . C lin . I n ve st. L u k a sh e v, M . E . , a n d We rb , Z . 1 9 9 8 . E C M sig n a lin g : o rc h e stra tin g
c e ll b e h a vio u r a n d m isb e h a vio u r. T re n d s C e ll B io l. 8, 4 3 7 4 4 1 . 103, 1 2 3 7 1 2 4 1 .
C e ll
7 0
S tra sse r, A . , H a rris, A . W. , B a th , M . L . , a n d C o ry, S . 1 9 9 0 . N o ve l
p rim itive lym p h o id tu m o u rs in d u c e d in tra n sg e n ic m ic e b y c o o p e ra -
tio n b e tw e e n myc a n d bcl-2. N a tu re 348, 3 3 1 3 3 3 .
S ym o n d s, H . , K ra ll, L . , R e m in g to n , L . , S a e n z-R o b le s, M . , L o w e , S . ,
J a c k s, T . , a n d Va n D yk e , T . 1 9 9 4 . p 5 3 -d e p e n d e n t a p o p to sis su p -
p re sse s tu m o r g ro w th a n d p ro g re ssio n in vivo . C e ll 78, 7 0 3 7 1 1 .
T h o rn b e rry, N . A . , a n d L a ze b n ik , Y. 1 9 9 8 . C a sp a se s: e n e m ie s w ith in .
S c ie n c e 281, 1 3 1 2 1 3 1 6 .
Va rn e r, J . A . , a n d C h e re sh , D . A . 1 9 9 6 . I n te g rin s a n d c a n c e r. C u rr.
O p in . C e ll B io l. 8, 7 2 4 7 3 0 .
Va u x , D . L . , C o ry, S . , a n d A d a m s, T . M . 1 9 8 8 . Bcl-2 p ro m o te s th e
su rviva l o f h e m a to p o ie tic c e lls a n d c o o p e ra te s w ith c-myc to im m o r-
ta lize p re -B c e lls. N a tu re 335, 4 4 0 4 4 2 .
Va ziri, H . , a n d B e n c h im o l, S . 1 9 9 8 . R e c o n stitu tio n o f te lo m e ra se
a c tivity in n o rm a l h u m a n c e lls le a d s to e lo n g a tio n o f te lo m e re s a n d
e x te n d e d re p lic a tive life sp a n . C u rr. B io l. 8, 2 7 9 2 8 2 .
Ve ik k o la , T . , a n d A lita lo , K . 1 9 9 9 . VE G F s, re c e p to rs a n d a n g io g e n -
e sis. S e m in . C a n c e r B io l. 9, 2 1 1 2 2 0 .
Vo lp e rt, O . V. , D a m e ro n , K . M . , a n d B o u c k , N . 1 9 9 7 . S e q u e n tia l d e -
ve lo p m e n t o f a n a n g io g e n ic p h e n o typ e b y h u m a n fib ro b la sts p ro -
g re ssin g to tu m o rig e n ic ity. O n c o g e n e 14, 1 4 9 5 1 5 0 2 .
We in b e rg , R . A . 1 9 9 5 . T h e re tin o b la sto m a p ro te in a n d c e ll c yc le
c o n tro l. C e ll 81, 3 2 3 3 3 0 .
We rb , Z . 1 9 9 7 . E C M a n d c e ll su rfa c e p ro te o lysis: re g u la tin g c e llu la r
e c o lo g y. C e ll 91, 4 3 9 4 4 2 .
Wh ite lo c k , J . M . , M u rd o c h , A . D . , I o zzo , R . V. , a n d U n d e rw o o d , P . A .
1 9 9 6 . T h e d e g ra d a tio n o f h u m a n e n d o th e lia l c e ll-d e rive d p e rle c a n
a n d re le a se o f b o u n d b a sic fib ro b la st g ro w th fa c to r b y stro m e lysin ,
c o lla g e n a se , p la sm in , a n d h e p a ra n a se s. J . B io l. C h e m . 271, 1 0 0 7 9
1 0 0 8 6 .
Wrig h t, W. E . , P e re ira -S m ith , O . M . , a n d S h a y, J . W. 1 9 8 9 . R e ve rsib le
c e llu la r se n e sc e n c e : im p lic a tio n s fo r im m o rta liza tio n o f n o rm a l h u -
m a n d ip lo id fib ro b la sts. M o l. C e ll. B io l. 9, 3 0 8 8 3 0 9 2 .
Wyllie , A . H . , K e rr, J . F . , a n d C u rrie , A . R . 1 9 8 0 . C e ll d e a th : th e sig n ifi-
c a n c e o f a p o p to sis. I n t. R e v. C yto l. 68, 2 5 1 3 0 6 .
Ya rd e n , Y. , a n d U llric h , A . 1 9 8 8 . E G F a n d e rb B 2 re c e p to r o ve re x -
p re ssio n in h u m a n tu m o rs. G ro w th fa c to r re c e p o r tyro sin e k in a se s.
A n n u . R e v. B io c h e m . 57, 4 4 3 4 7 8 .
Z h u , J . , Wa n g , H . , B ish o p , J . M . , a n d B la c k b u rn , E . H . 1 9 9 9 . T e lo -
m e ra se e x te n d s th e life sp a n o f viru s-tra n sfo rm e d h u m a n c e lls w ith -
o u t n e t te lo m e re le n g th e n in g . P ro c . N a tl. A c a d . S c i. U S A 96, 3 7 2 3
3 7 2 8 .
Z u o , L . , We g e r, J . , Ya n g , Q . , G o ld ste in , A . M . , T u c k e r, M . A . , Wa lk e r,
G . J . , H a yw a rd , N . , a n d D ra c o p o li, N . C . 1 9 9 6 . G e rm lin e m u ta tio n s
in th e p 1 6
I N K 4 A
b in d in g d o m a in o f C D K 4 in fa m ilia l m e la n o m a . N a t.
G e n e t. 12, 9 7 9 9 .