Conf Munique 2012 - Professor Serge Jurasunas

Target Therapies
P53
BCL-2 Apoptosis
Survivin
Angiogenesis
Telomerase
Resistance
Molecular Targets P21
strategies Dissemination
COX2
VEGF
IL 6
Survival
2nd International Conference
Proliferation
NF.KB on Complementary Oncology
Primary and compensatory Organized by the German Society of Oncology
pathway 15th-17th June 2012 – Munich - Germany
Speaker: Serge Jurasunas M.D. (Hom) N.D.

Member of the Society of Integrative Oncology (USA)
Topic: How to treat cancer by targeting P53, BCL-2, Survivin:

Prevention, diagnostics, targeting therapies – Case studies.
1
Holiterapias
Integrative and Complementary Medicine
Integrative Medicine – Complementary Oncology
Our work focus 3 directions:
● Cancer prevention (earlier diagnosis)
● Treatment of cancer
● Prevention of disease recurrence
Complementary Test: Live blood analysis, thermography, electromagnetic

field test, oxidative stress tests, apoptosis markers.
Treatment: immunomodulation, angiogenic therapy, antioxidant therapy, stem

cells, anticancer vaccin, orthomolecular medicine, detox, anticancer diet, SGE
therapy. 2
Outline of the lecture
● What is cancer ?
● Cancer a matter of life or death.
● Molecular markers P53, BAX, BCL2, Survivin, P21 and their proteins
involved in cancer initiation, progression, drug resistance, serve as a new
diagnostic, prognosis, and response to treatment.
● Clinical application of molecular markers and their proteins with cancer

patients.
● Dietary foods and compounds to target gene expression.
Suggesting reading:
3
Pierre Hainaut and Kas G. Wiman – P53- 25 years of research – Edit Springer – 2007
What is cancer – A matter of life and dead
● Cancer is an accumulation of
abnormal cells which divide
without control, evade apoptosis,
accumulate mutations and are
able to grow and invade other
tissues.
● Cancer cell survival is totally

dependent on defective P53 a key
hallmark of cancer and excess of
anti-apoptotic proteins for its
survival, growth and invasion.
● The balance between pro-

apoptotic signals and survival
signals determine the cellular
fate.
Evan G. littlewood T.: A matter of life and cell death – Science 1998: 281 – 1317-1322 4
Hallmark of cancer (Dr. Hanahan and R. Weenburg)
● Several basis elements involved in the malignant process
■ For a population of cells to exhibit

uncontrolled growth, abnormal cell
proliferation is necessary among
other things, that cells must escape
the induction of apoptosis.
■ Apoptose is a necessary
development mechanism a hallmark
of cancer to act as a barrier against
cancer and associate with most
hallmarks
5
Tuning in on the P53
DNA Damage and
Tumor Suppression
Programs
6
Apoptotic pathway
7
P53 and Cancer
P53 is the “Guardian of the genome” which instructs

genetically damaged cells to repair or die.
Thus, P53 guards against changes to cells that might lead to

tumor formation and cancer.
P53 mutation or inactivatition appears necessary to develop

many forms of cancer.
P53 is mutated in about half of all tumors.
Alex Sigal and Varda Ratter – Oncogenic Mutations of the P53 Tumor suppressor: The 8
demons of the Guardian of the Genome – Cancer Res. Dec. 12.200-60-6788
P53 mutation distribution in cancer
P53 gene is one of the most

commently mutated gene in 50% -
60% of all cancer.
Brain – 50%
Breast - 20 to 45%
Ovary – 40% to 60%
Colon - 40 to 70%
Lung – 70%
Melanoma – 50%
9
Oncogenic function of mutant P53 proteins
Tumor initiation – progression - invasion
Gain of Function (G.O.F)
Mutagenesis Dominant negative effects
Transcriptional activation is
impaired – Mutant P53 protein
fails to bind specific DNA
Loss of wildtype P53 Mutant P53 sequence.
function
Pro-apoptotic signals via BH3 only

Targeting proteins are impaired (BAX, BAK,
anticancer Puma, Noxa)
therapy
Accelerate tumor progression
Promote angiogenesis
Invasion Activation of specific target genes EGFR/1,
Metastasis RAS, interference with the TGFB growth arrest
Enhanced resistance to
Immunosupression control pathway.
chemotherapy
Interference (boost) with NF.KB
Downregulation of the E-Cadherin cell-cell
addhesion molecule enhances motility.
A key event to E.M.T. in tumor progression
ID4 – E.2F.1 – Promotes angiogenesis
Patricia A.J. Muller, Karen H. Vonsden and Jim C. Norman – P53 and its mutants in 10
tumor cell migration and invasion – JCB Home – January 2011 – Vol. 2 209-218
Mutation that lead to EGFR overexpression (upregulation) or
overactivity have been associated with a number of cancers. 11
E-Cadherin associate with invasive cancer
■ One half of the invasive ductal

carcinoma that develope distant
metastases have aberrant –
E-cadherin protein expression.
■ Increasing evidence that

cancer cells may re-express E-
cadherin protein once they reach
distant sites.
■ Distant metastases express

E-cadherin often more strongly
than the primary tumor.
Paul J. Kowaski, Mark A., Ruben, Celina G. Kleen – E-Cadherin expression in primary carcinoma of
the breast and its distant metastases. – Breast Cancer Res 2003 – 5 R. 217 - 222 12
SOD NF-KB is a redox sensitive transcription
factor coordinate regulators of immunity
stressors inflammation, development, cell
proliferation and survival
ROS
cytokines Cell
ROS high concentration
Survival
Caspases inhibitors
BCL2 (elevated expression)
response P53
IK-BKinase
enhance
Sequestred and inactivated
NFKB Cytosol Release NFKB translocates to the nucleus and

bind to DNA at the promotor region and
activates the transcription of target genes.
Suppression
SOD
ROS low concentration
Modulate cell death/survival Dead cell
13
Serge Jurasunas 2008
Cooperation Response Genes – P53 + RAS activation
Oncogenic mutations in P53 and protein RAS have individually limited effects on promotion of
cancer. However they cooperate to transform normal cells into cancer cells and accelerate
tumor growth.
P53 and RAS mutated in the same cells they synergistically regulate a subset of genes (AB)
know as “Cooperation Response Genes” (CRG’s) mediator of tumor formation. 14
Hallmark of P53 Mutation
EGFR/Integrin
Angiogenesis
Metastasis Invasion
Anti-apoptotic proliferation
survival
E.2F1 RAS
Promote Angiogenesis
angiogenesis Proliferation
Invasion
Decreased apoptosis
ID 4 C-Myc
Promote angiogenesis Decreased apoptosis
Tumor proliferation Proliferation
Promote malignant
progression
P53
Immunosuppression
Antiapoptotic Mutation
Inflammation
Angiogenesis E-Cadherin
NF.KB Metastasis Associate with
E.M.T.
Promote invasiveness
Immunosuppression
and metastases
enhance motility
Increase evasion
angiogenesis
TGF. Beta
15
Copyright Serge Jurasunas 2012
16
P53 mutation –
Causative factors in the progression of cancer
Conclusion:
Overall
■ Associated with increasing cancer cell resistance to chemotherapy or

other treatments that trigger apoptosis.
■ Associated to metastatic potential, faster tumor growth and
invasiness.
■ Associated with aggressive cancers such as breast, Ovarian and
worst of all, Survival.
Sood A.K., J.I. Dolan M. et al (1999) – Distant metastases in ovarian cancer Association
with P53 mutations – Clin Res. 5 – 2485-2490. 17
P53 – Potential biomarker: Cancer strategy
■ Potential biomarker
■ Earlier detection
■ Diagnostic Anticancer Strategy

■ Prognostic
■ Relapse
■ Response to treatment
■ Re-establish chemo sensitivity
Haupt S., haupt Y. (2004) – improving cancer therapy through P53 management – Cell cycle: 3:
912-916 18
Tumor – Invasion - Metastasis
Several basic elements are involved in the malignant process
■ Evading apoptosis
Mutated P53
■ Elevated NF.KB activity
■ Overexpressed of BCL2
■ Activated Pi3 – Kinase pathway
■ Loss of E-Cadherin
■ Substained angiogenesis
■ Overexpressed Survivin
90% of human cancer deaths are caused by metastases

19
A new anticancer treatment
Therapy to target Mutant P53
Reactivating Mutant P53 using small molecules
Enhance drugs
sensitivity
Reverse
Mutant P53 Wild type
Gain of function Normal function Tumor

Increasing apoptosis regression
Self-destruction of
cancer cells
Tumor growth
Evasion of
Angiogenesis
Metastases
Andrea Ventura, david G. Kirch, Margaret E., Mc Lughlin, David A. Tuveson, Laura Lintault, 20
Tyler Jacks – Restoration of P53 function leads to tumor regression in vivo – Nature 445 – 661-
665 (Feb.2007)
Targeting BCL-2, BAX, Survivin and other
proteins involved in apoptosis as a mean to
modulate drug resistance.
Tito Fojo and Susan Bates – Strategies for reversing drug resistance –
21
Oncogene (2003) 7512 - 7523
P53 transcriptionaly control BCL-2 and BAX to
mediate apoptosis
22
BCL-2 – Anti-apoptotic proteins
■ BCL-2 enhances angiogenesis

and promotes resistance of cancer
cells to a wide range of anticancer
agents and immune cells.
■ BCL-2 overexpression is
associated with metastasis shorter-
disease-free survival in a variety of
cancers.
■ Down-regulation of anti-apoptotic
BCl-2 members sensitize cells to
chemotherapy.
Donatello Del Bufalo, Annamaria Biraccio, Carlo Leonetti and Gabriella Zupi – BCL-2 23
overexpression enhances the metastatic potential of human breast cancer line FASE B –
950 – Vol.11 – Oct. 1997.
BCL-2 – block apoptosis and increase cancer cells resistance
A layer of pink normal cells is shown

in the background and a mass of green
tumor cells is in the foreground. Small
yellow molecules representing
chemotherapeutic drugs are present.
The tumor cells are not undergoing
apoptosis in response to the
chemotherapy.
Pink – normal cells

Green – Mass of tumor cells
Small yellow molecules – chemotherapeutic drugs
Tumor cells are not undergoing apoptosis 24

Pro-apoptotic BAX proteins – A tumor suppressor
inactivated in many cancer.
A key player in cell death induced by anticancer drugs
■ Bax gene expression is inactive in approximately one-third of invasive

breast cancer (specimen of 150 patients) (1)
■ Another study in 119 women with metastatic breast cancer, patients whose
tumor has lost Bax activity had poor responses to chemotherapy, faster time
to progression and shorter overall resistance.
■ Enhanced expression of Bax proteins is associated with good response to

chemotherapy/radiation in vivo.
(1) John Red – Bax gene expression in breast cancer – The Burknam Institute for Medical
Research. Research project Award – Calif. Breast Cancer research program – 1995.
(2) Krajewski S., Blomquist C., Fransilla K. et al – Reduced expression of proapoptotic gene
BAX is associate with poor response rates to combination chemotherapy and shorter 25
survival in women with metastatic breast adenocarcinoma – Cancer Tes. 1995 – 55: 4471-
4478
Induction of the mitochondrial apoptotic pathway
by cisplatin
- Activates the proapoptotic

protein BAX
- release of cytochrome C
- induce the formation of the

apoptosome
- APAF1 – cytochrome C complex
- Activation of procaspase-9
leading to activation of caspase-3
– cell death
26
Predictive value of BAX/BCL2 in
chemo/radiation regimen
Cells that overexpress BAX enhance cell death.
Cells that overexpress BCL-2 reduce the suceptibility to apoptosis.
Because BAX proteins antagonize BCL-2’s anti-apoptotic function, it

is likely that BCL-2/BAX ratio determines the suceptibility of a cell to
apoptosis and determines the therapeutic response of
chemo/radiation regimen to apoptosis stimuli.
Scopa, Chriscula, Vagionas, Constantine, Kardimaki, Dimitris, Kourelis, Athanasias C. –

BCL-2/BAX ratio as a predictive Marker for Therapeutic Response to radiotherapy in patient
with rectal cancer – Immunotochemistry x Molecular Morphology – Dec.2001 – Vol.9 – Issue 27
4 – 329-334
The involvement of BAX in zinc induced mitochondria
apoptogenesis in malignant prostate cells
The effects of zinc on Bax and Bcl-2 levels of

mitochondria isolated from PC-3 cells that were The effect of 15 μM zinc of PC-3 cells on the total
exposed to medium supplemented with zinc. The cells cellular levels of BAX and BCL-2.
were treated with medium supplemented with 15 μM
zinc for 180 minutes versus no zinc treatment. BAX after three hours of zinc treatment.
After six hours BAX level increased by 180%.
By 30 minutes exposure to zinc, MT level of BAX was In contrast BCL-2 increase of about 30% in nine
increased by about 100% - by 260% in 180 minutes. hours of zinc treatment.
Zinc treatment caused a much lower increase in BCL-2 Zinc treatment cause the relative BAX/BCL-2 ratio to
– about 30% in 30mn and 80% by 180 mn. increase – pre-treatment ratio of 1/1 or 2.5/1 after
Zinc treatment resulted in a 2-fold increase in the treatment.
BAX/BCL-2 ratio
28
Pei Feng and all – The involvement of BAX in zinc-induced Mitochondrial Apoptogenesis in Malignant
Prostate cells – Molecular Cancer 2008 – 7.25
Survivin – A new inhibitor of Apoptosis
Survivin is an inhibitor of apoptotic proteins (IAP’s) inhibit Caspase 3,

Caspase 7, Caspase 9.
Survivin expression is absent normal tissue however overexpressed

in a wide spectrum of tumors: Ovary, colon, prostate, neuroblastoma,
breast and associate with strong drugs resistance and toxic adverse
effects to a wide spectrum of anticancer agents.
WT P53 may transcriptionaly repress Survivin gene expression P53

mutation is associate with dramatic expression of Survivin.
Extracellular Survivin released is promptly captured by neighboring

cancer cells, enhancing their proliferation, invasive capacity and
resistance to therapy.
29
Emerging modalities to treat cancer patients
Survivin pools
■ Associate with different compartimentalization of the proteins in the
cells.
■ Nuclear cytoplasmic and mitochondrial pools of Survivin (Fortugno

et al 2002).
■ Nuclear localization is linked with the capacity of Survivin to

regulate cell division whereas MT survivin is associated with
protection from apoptosis.
■ Extracellular Survivin released is promptly

captured by neighboring cancer cells, enhancing
their proliferation, invasive capacity and resistance
to therapy (Mera et al 2000 Klan et al 2011)
■ T-cells mount a vigourous cytotoxic response to

survivin peptides in vitro and in vivo.
Anderson, MH et al (2001) – Identification of a cytotoxic T-

Lymphocyte response to the apoptosis inhibitor protein survivin 30
in cancer patients – Cancer Res. 61-869-872.
Function of survivin as inhibitor of apoptosis
Dobi T., Beltrami E., Wall N.R., Plescia J., Altieri D.C. – Mitochondrial survivin inhibits 31
apoptosis and promotes tumorigenesis – J. Clin invest. 2004: 114-1117 – 27.
Survivin plays a critical role in Angiogenesis
■ Survivin is strongly up-regulated in angiogenically stimulated

endothelium in vitro, in vivo increasing drug resistance.
■ increased Survivin expression protected E.C. from Apoptosis during the

proliferative and remodeling phases of angiogenesis.
■ Targeting Survivin in E.C. may be a potential strategy to inhibit – tumor

– associated vessel growth (O’Connor et al 2000).
Trans J. Master, Z. Yu, Il et al – A role for Survivin in chemoresistance of endothelial cells

mediated by VEGF – Proc. Natl Acad. Sci. USA 2002: 99-4349-54.
Florence Hofman: Survivin target for breast cancer brain metastases: Breast Research Cancer –
Prog. Uni Southern Calif. 2005.
S.P.T.X.H. Jiang, M.C. M. Lin J.T. Cui et al. – Supression of Survivin expression inhibits in vivo
Tumorigenicity and Angiogenesis in Gastric Cancer. – Cancer Res. – November 15, 2003: 63 (22) 32
7724 - 7732
Angiogenesis: Cascade of Events
33
Tumor – Associated Angiogenesis
Targeting mutant P53

proteins and Survivin
overexpression may be
an effective combination
therapy to inhibit
angiogenesis in
advanced breast and
brain tumors.
Florence Hofman: Survivin for breast cancer brain metastases – University of Southern
Calif. Award cycle : 2005 (cycle 11) 34
New approach to tumor growth and metastases
1 – Attacking tumor chemotherapy
2 – attacking its vascularazation cytotoxic T-cell response
against Survivin protein Activation of apoptosis in

tumor cells
Less resistance
Xiang R., Mizutani N., Luco Y., Chiodoni C., Zhon H., Mizutai M. Bcker J.C. (January 2005)
DNA vaccine targeting
35
Angiogenesis
A complex process - I
Cells deprived of oxygen Inflammatory
(white) cells
emit angiogenic signals
Red
blood
VEGF cells
Signal source :
Tumor cells VEGF
Hypoxia
Inflammation VEGF
Basement
Endothelial membrane
cells
www.sergejurasunas.com
Angiogenesis
A complex process - II
Inflammatory
(white) cells
Red
Signal source blood
Migration Proliferation cells
MMP action Basement

membrane
Endothelial
cells
Angiogenesis
A complex process - III
A new blood vessel is formed
Functional vascularization
Oxygen and nutrient
supply
Clinical application of L.C.E. at Holiterapias Institute with
successful response in cancer patient during a 12 years
period.
This include a wide variety of different cancer types such: prostate, multiple
myeloma, lung, colorectal, breast, ovaries, melanoma pancreas.
-Benefit survival extension.
-Significative reduction of tumor size.
-Decrease of metastasis specially bone metastasis.
-Elimination of secondary nodules.
-Our patients take L.C.E. during several years without

toxicity.
-Suggested in anti-cancer dormancy therapy
39
Example of tumor vascularization in a case of advanced
inoperable breast cancer without conventional treatment
Observation: Imaging infrared thermographic camera system
Tumor, large inflammation and Elimination of the vascularization

vascularization angiogenic. Tumor remains inactive.
Photo nº 1 Photo nº 2
Duration of the treatment: 7 months

No surgery or conventional therapy 40
LCE + immunomodulator Jurasunas Serge, ND
Ribozyme – mediated inhibitor of survivin expression
sentitizes prostate cancer cells to cisplatin – induced
apoptosis
1 – Cisplatin treated cells in

the absence of the caspase
inhibitor (Grey)
2 – Cisplatin treated cells

with the presence of the
caspase inhibitor
Marzia Pennati et al – Ribozyme – mediated inhibition of survivin expression increases spontaneous

and drug – induced apoptosis and decreases the tumorigenic potential of human prostate cancer41
cells – Oncogene (2004) 23: 386-394
Increasing Survivin Expression in different grades of
prostate carcinoma
Associated with aggressive prostate cancer
100%
80%
60%
40%
20%
0%
Normal prostate Primary grade Primary high High grade
tissue prostate grade prostate prostate
carcinoma carcinoma carcinoma
Ashfoq R., et al – Survivin is associate with feature of biologically aggressive prostate

carcinoma – Cancer – 2004 – 100 – 751 - 757 42
Overexpressed Survivin and prostate cancer
■ Study from a collection of prostate cancer and normal tissue in chinese

cohort (1)
1 – Survivin protein is virtually undetectable in normal prostate

tissue
2 – Survivin is highly expressed in prostate cancer
Overall:
Survivin expressed in 24 of 28 (85.7%) prostate cancer specimens

No survivin expressed detected in 22 normal prostate tissues
■ Difference is statistically significant (PL 0,001)
■ Strong correlation between Gleason Score and Survivin expression among

studied cases. (R= 0,7446) PL0,0001
(1) Xichum et al: (Survivin gene silencing sensitizes prostate cancer and cells to selenium
growth inhibitor. BMC – Cancer – 2010-10-418 43
Survivin in breast cancer
Study done at St. Vincent’s University Hospital
University College Dublin (Irland) 2004
120
90
■ Survivin is virtually absent from
60 normal tissues.
30 Very high levels in malignant breast
0
tumors.
Normal Cancer Metastases
■ Study of 500 breast cancer patients:

Patients with high levels of survivin: reduced survival time and more subject to
disease recurrence.
■ Strong predictor of poor prognosis
Hinnis A.R. Luckett J.C. Walker R.A. – Survivin is an independent predictor of short-term 44
survival in poor prognostic breast cancer patients – BR. J. Cancer 2007 – 96-639-45
Survivin: a new biomarker and emerging as a
attractive target to cancer
■ Increased survivin expression is associated with lymph

nodes, invasion, metastases risk of recurrence and
decreased overall survival in several malignancies.
■ Survivin detection can serve as an early marker of cancer,

for detecting metastases, breast cancer and prostate
cancer recurrence.
Yong-Gang L.V. – The role of survivin in diagnosis, prognosis and treatment of breast cancer.
J.Thorac Dis. 2010 – 2: 100 - 110 45
P21 – The second line of cellular anticancer defense
(Tumor suppressor)
● P21 – WAF1/cpF is a potent cyclin-dependent Kinase inhibitor, key

mediator of P53 dependent or independent cell cycle arrest
● P21 could self-activate independently to a non-active P53, induce

apoptosis and promote self-destruction of damaged cells as a response
to toxic agents and radiation.
● Several reports suggested that P21 and Survivin are also functionally
associated.
● Conversely: Mutation of P53 gene expression in ovarian cancer is

usually associated with decreased P21 gene expression
Elbendary A.A., Cirisnano F.D., Evan A.C., Davis et al. – Relationship between P21 expression and
mutation of P53 tumor suppressor gene in normal and malignant ovarian Epithelial cells – Clin. Cancer
Res. – 2.1571-1575
Komiya T. et al. 46
Wong Yonli et al – The expressions of Survivin Protein and the Relationship between P21 (WAF1) and
P53 protein in infiltration ductal cancinoma on breast – Journal of Basic Clinical Oncology 2006/04.
Correlation between P21 gene expression and P53
mutation in normal and malignant epithelial cells
■ Study: 23 primary ovarian cancer.
● Normal levels of P21 RNA were seen in 4/7 (57%) cancer with wild
type P53.
14/16 (88%) cancer with mutant P53 had reduced P21 expression
(P<0,005).
● Irradiation of normal and malignant ovarian epithelial cells with wild

type (but not with mutant) P53 resulted in induction of P21.
Conclusion: Induction of P21 is a feature of P53 – mediated growth inhibition

in normal ovarian epithelial cells.
Conversely: Mutation of P53 gene expression in ovarian cancer usually is

associate with decreased P21 gene expression.
Brugaroles J. et al – Radiation induced cell cycle arrest compromised by P21 deficiency – Nature
1995: 377-552-557 47
P21 in cancer prognosis
■ Loss of P21 is associated with development of

metastasis, poor prognosis and decreased survival
in certain cancers: colorectal, gastric, ovarian,
breast.
Zirbes T.K. et al – Prognostic impact of P21/WAF1/Cp1 in colorectal cancer – Int. J. Cancer – 2009:
89: 14-18
Komiya T. et al – P21 expression as a predictor for favorable prognosis in squamons cell
carcinoma of the lung – Clin.Cancer Res. 1997: 3.1831-1837 48
49
Cancer patients doing the genes tests
and their proteins
Nurse table
Venous blood
Consultation
Laboratory
Method: Elisa Assay
Quantitative Polymer Chain reaction (P.C.R.)
50
Example of P53 assay -
two cases of breast cancer remission
F.43 years old F. 48 years old (April 2012)

Test done 1 year after remission Test done 3 months after remission
P53 Gene Expression P53 Gene Expression

1.3x106 copies/ml of plasma Not detectable
(100.000) Reference range: 10-50 copies
Reference range: = 1000 copies
P53 Protein Level
P53 Protein Level 0.02 units of mutated protein/ml of
5.2 µ of normal protein/ml of plasma plasma
Reference range: 0.33 copies Reference range: 0.10 – 1.00 units
W.T
Not detectable
Reference range: 0.02 units
Self-destruction of abnormal/stressed
cells The tumor suppressor gene is not active
and produced only a trace of P53
mutation -stressed/transformed cells are
not self-destructed.
Risk of recurrence 51
F – 44 years – non cancerous patient – 2009 (Stressed cells)
P53 Assay
P53 Gene Expression P53 Protein Level

18.788 copies/ml of plasma 57.3 µ of normal protein/ml of plasma
Reference range Reference range
50 copies 0.33 units
Comment:
There is a population of stressed cells with damaged DNA as it is indicated by
P53 gene expression and high level of WT protein.
Cancer lesions are unlikely because of the absence of mutated P53 protein .
The cancer defense is functioning.
52
Patient F – 62 years old
Breast cancer recurrence with distant metastases to lung and liver
P53 Protein Level P53 Protein Level

P53 Gene Expression mutated (Wild /normal)
Not detectable Not detectable Not detectable
10-50 units 0,02 units
BCL2 Gene Expression

Not detectable Poor prognosis
Reference range
10 units
Survivin Gene Expression P21 Gene Expression

7.059 units/ml of plasma 2.543 units/ml of plasma Ratio 0,36
Reference range dominance Reference range
10 units 10-50 units
Comments: Poor prognosis because of P53 inactivation and very high activity of Survivin
gene expression. Only a small fraction of cancer cells self destruct. The patient didn’t
respond to various line of chemotherapy regimen: 5 Fu cytofosfamide, pacitaxel, Herceptin, 53
etc., increasing toxic effects-short survival.
Patient F. with a 4 years complete remission
of pancreatic cancer (Townsend Letter – August/Sept 2009 USA)
P53 Test
1st Test – 4.5.2009 2nd Test – 4.8.2009 3rd Test – 17.11.2009
P53 Gene Expression P53 Gene Expression P53 Gene Expression

65.000 copies/ml of plasma 8.9 x 108 copies/ml of plasma 1.2 x 106 copies/ml of plasma
Reference range: 1000 copies Reference range: 1000 copies Reference range: 1000 copies
P53 Protein Level P53 Protein Level P53 Protein Level

4.5 µ of abnormal (mutated) 1.5 µ of normal protein/ml of 67.4 µ of normal protein/ml of
protein/ml of plasma plasma plasma
Reference range: 0.33 copies Reference range: 0.33 copies
P53 Wild Type
Indetectable
Reference range: 0.33 copies
We reversed mutant P53 to a Now the P53 gene is still active

wild type but the actived gene and produces high a level of
produced normal P53 protein P53 normal protein leading to
only to some extent. Cancer increased self-destruction of
cells are not fully self- cancer cells.
destroyed Tumor regression can be
anticipated
54
F – 69 years – pancreatic cancer – no metastasis
(chemo+dietary agents before) 2012

P53 Gene Expression mutated Wild normal)
Not detectable 5.60 units/ml of plasma Not detectable
10-50 units 0.10 – 1.00 units
BCL2 Gene Expression Anti and pro- tumor activities

85 units/ml of plasma
Reference range
10 units Growth of the tumor
mass is limited.
Self-destruction of
Survivin Gene Expression P21 Gene Expression cancer cells
Not detectable 7.624 units/ml of plasma
dominance 55
Patient F - 49 years old – ovarian cancer –
stage III - recurrence
Recurrence in 2009
1.7.2009 18.1.2011 (remission)

3.285 copies/ml of plasma 1.2 x 108 copies/ml of plasma
Reference range:50 copies Reference range:50 copies

normal normal
Not detectable 125.0 µ of normal protein/ml of plasma
Reference range: 0,33 units/ml of plasma Reference range: 0,33 units/ml of plasma
P53 Protein Level Comments:

mutated The treatment reversed the P53 mutant
15.75 µ of abnormal protein/ml of plasma with oncogenic function. The tumor
Reference range: Not detectable
suppressor gene is now very active with
an increased level of normal proteins.
Comments: Active process of self-destruction of
Cells are resistant to destructive cancer cells leading to tumor regression. 56
agents that trigger apoptosis.
Patient M – 81 years old – recurrence colon cancer
- Liver metastasis
1st Test: 9/3/2011 2nd Test: 11/8/2011

Reference range:
Not detectable 10-50 units/ml of plasma 1.180 units/ml of plasma
P53 Protein Level P53 Protein Level n

normal Reference range: normal
Not detectable 0,33 units/ml of plasma Not detectable

mutated mutated
10.88 units/ml of plasma Not detectable
Reference range:
BCL2 Gene Expression BCL2 Gene Expression
10 units
340 units/ml of plasma apoptosis Not detectable
Reference range:
BAX Gene Expression BAX Gene Expression
2 pools of 10-100 units
Not detectable resistance 409 units/ml of plasma
Reference range:
Survivin Gene Expression Survivin Gene Expression
10 units
129 units/ml of plasma Not detectable
Reference range:
P21 Gene Expression 10-50 units P21 Gene Expression 57
Not detectable Not detectable
Patient M – 50 years old – lung cancer –
Molecular Marker Test 09.05.2011
Before chemotherapy

Reference range Apoptosis is not functioning Reference range
10-50 units 0,10 – 1.00 units
BCL2 Gene Expression BAX Gene Expression

340 units/ml of plasma 303 units/ml of plasma Ratio 0,89
Reference range Pre cancerous cells Reference range
10 units transformed into 10-100 units Malignant potential limited
malignant ones due to anti-tumor activities
of BAX - P21 genes
1.028 units/ml of plasma 552 units/ml of plasma Ratio 0,53
Possible result from
chemotherapy
dominance 58
Poor prognostic
Chemotherapy and support of complementary oncology
Result of the Hospital Scan (18.07.2011) after chemotherapy coupled with

our complementary approach.
In comparaison of the past Scan done in April 2011 we observed a

significative reduction of lesion size in the right lobe.
1 – 2.3 x 1.4 cm 3.8 x 2.5 cm
2 – 2 x 2.1 cm 2.8 x 3.1 cm
3 – 1.6 x 1.4 cm 3.4 x 3.7 cm
4 – 2.4 x 1.5 cm 5.8 x 3.6 cm
Complementary approach – Targeting angiogenesis – immunostimulation

59
The patient feels well without side – effects from chemotherapy
During chemotherapy/radiation
3units/ml of plasma Not detectable 0.1 units/ml of plasma
Reference range Apoptosis is not functioning Reference range
10-50 units 0,10 – 1.00 units
Start functioning
Decrease
Reference range Before 340 units Reference range
Increasing activity
5 units/ml of plasma 4.527 units/ml of plasma Ratio 905,4
Reference range Before 1.028 units Before 552 units
10 units
60
dominance
Case of a patient 71 years old with a advanced prostate cancer
Gleason (5+4) – Stage
-Multiple bone lesions
-Very high P.S.A. Level
-Very severe anemia
-Poor physical condition (lost 25 Kg)
-No surgery or conventional treatment
Ashfaq R.: Survivin expression is associate with features of biologically aggressive 61

prostate carcinoma – Cancer 2004 – 100 – 751 - 757
PSA decrease over 5 months from 680,400 ng/ml to 89.00 ng/ml
No Chemotherapy
700
600
500
400
P.S.A.
300
200
100
0
02,11,2011 17,11,2011 17,12,2011 19,01,2012 02,02,2012 22,02,2012 6,03,2012 9.04.2012
62
M – 71 years – advanced prostate cancer –
Molecular Marker Test 17.12. 2011

10-50 units 0,10 – 1,00 units

Protumoral
60 units/ml of plasma Not detectable dominance
Pool of
resistance

24.210 units/ml of plasma 12.840 units/ml of plasma Ratio 0,53
Reference range Pool of Reference range
10 units resistance 10-50 units
dominance 63
M – 71 years – advanced prostate cancer –

4 units/ml of plasma Not detectable 0,02 units/ml of plasma
10-50 units Should be more active 0,10 – 1,00 units

10 units 10-100 units Active process of the destruction
(apoptosis) of the tumor mass
from the applied treatment
Anticancer dominance
dominance 64
Patient 62 years – cancer of bladder, ureter and prostate
6th February 2012 12th March 2012 16th May 2012

Not detectable 149 units/ml of plasma 180 units/ml of plasma
Reference range: 10 - 50 copies Reference range: 10 - 50 copies Reference range: 10 - 50 copies
P53 Protein Level (mutated) P53 Protein Level (mutated) P53 Protein Level (mutated)
2.8 µ of abnormal protein/ml of Not detectable Not detectable
plasma Reference range: Not detectable Reference range: Not detectable
Reference range: Not detectable
P53 Wild Type P53 Wild Type P53 Wild Type

(normal) (normal) (normal)
Not detectable Not detectable 1.3 units/ml of plasma
Reference range: 0.10 - 1.00 units Reference range: 0.10 - 1.00 units Reference range: 0.10 - 1.00 units
65
Patient 62 years – cancer of bladder, ureter and prostate
6th February 2012 12th March 2012 16th May 2012
BCL-2 Gene Expression BCL-2 Gene Expression BCL-2 Gene Expression

Not detectable 142 units/ml of plasma Not detectable
Reference range: 10 units Reference range: 10 units Reference range: 10 units
Ratio
0,95
BAX Gene Expression BAX Gene Expression BAX Gene Expression
Not detectable 136 units/ml of plasma Not detectable
Reference range: 10 – 100 units Reference range: 10 – 100 units Reference range: 10 – 100 units
Survivin Gene Expression Survivin Gene Expression Survivin Gene Expression

Reference range: 10 units Reference range: 10 units Reference range: 10 units
Ratio Ratio
1,12 6,5
Reference range: 10 - 50 units Reference range: 10 - 50 units Reference range: 10 - 50 units
Good response to chemotherapy End of May 2012

No adverse effects Scan- Biopsy-Stop chemotherapy
No tumor – No metastasis 66
Patient M. 68 years – possible prostate cancer
1st Test 2nd Test 3rd Test – 1st June

1.486 units/ml of plasma 89 units/ml of plasma 45 units/ml of plasma
Reference range: 10 - 50 copies Reference range: 10 - 50 copies Reference range: 10 - 50
copies
P53 Protein Level (mutated) P53 Protein Level (mutated) P53 Protein Level
Not detectable Not detectable (mutated)
N.D.
Reference range: Not detectable Reference range: Not detectable
Reference range: Not
detectable
P53 Wild Type P53 Wild Type P53 Wild Type

(normal) (normal) (normal)
Not detectable 0.05 units/ml of plasma N.D.
Reference range: 0.10 - 1.00 units Reference range: 0.10 - 1.00 units Reference range: 0.10 -
1.00 units
67
Patient M. 68 years – possible prostate cancer
1st Test 2nd Test 3rd Test
BCL-2 Gene Expression BCL-2 Gene Expression BCL-2 Gene

Not detectable 94 units/ml of plasma Expression
Reference range: 10 units Reference range: 10 units
Reference range: 10 units
Ratio
BAX Gene Expression BAX Gene Expression 1,0 BAX Gene Expression
81.699 units/ml of plasma 94 units/ml of plasma N.D.
Reference range: 10 – 100 units Reference range: Reference range: 10 – 100
units
10 – 100 units
Survivin Gene Expression Survivin Gene
51.548 units/ml of plasma Survivin Gene Expression Expression
Reference range: 10 units 105 units/ml of plasma 30 units/ml of plasma
Reference range: 10 units Reference range: 10 units
Ratio
P21 Gene Expression 1,26
Not detectable P21 Gene Expression P21 Gene Expression
Reference range: 132 units/ml of plasma N.D.
10 - 50 units Reference range: 10 - 50
Reference range: 10 - 50 units
units
TM2 – Pyruvate Kinase TM2 – Pyruvate Kinase

142 units/ml of plasma 49,2 units/ml of plasma
Reference range: 5.0 – 15.0
Reference range: 5.0 – 15.0 units
units
Decrease
68
F – 44 years – Breast cancer Stage III with
multiple large lesions (30) to liver
Test done before chemotherapy (April 2012)

200 units/ml of plasma Not detectable Not detectable
10-50 units 0.10-1.00 units
Should be more actived
BCL-2 – major factor leading to
disease progression.
8 x 106 units/ml of plasma 167 units/ml of plasma Ratio 0,02
10 units 10-100 units Vascular Endothelium
Growth Factor
Pool of high resistance
Reference range 10-100 units

69
dominance Bad prognosis
F – 44 years – Breast cancer with metastases – Test done
during chemotherapy (May 2012)
The Anti-tumor effect of the Applied treatment

427 units/ml of plasma Not detectable 0.4 units/ml of plasma
10-50 units 0.10-1.00 units

10 units 10-100 units Vascular Endothelium
Growth Factor
Not detectable
Reference range 10-100 units
70
dominance Bad prognosis
71
Chemotherapy: The patient feels well, no adverse effect – tumor
reduced by end of May: Tumor decrease, diminution of the axillary
lymph nodes: surgery possible - Karnofsky score: 90
The current pattern has become more anti-tumor compared with the
previous pattern.
Chemotherapy increasing efficiency
P53 gene expression WT protein level increasing apoptosis pathway

Increasing
BAX gene expression increasing activity increasing cancer cells destruction

activity
BCL-2 decreasing activity (8.000.000 – 769 units) less resistance from cancer cells
Survivin
Same ratio although P21 is increasing better response to chemotherapy
P21
VEGF – shot down (2353 units – ND) less resistance (decreasing angiogenesis)
72
The treatment that successfuly targeted the Molecular
Markers, specially high BCL2 overexpression, VEGF
and decrease tumor size
Liquid Cartilage Extract (L.C.E.) – (frozen liquid)

One ampoule per day
Oligopeptide (S. Jurasunas product)

12 tablets per day (1)
Curcumin (target P53, NF.KB – angiogenesis – immune cells)

8 gr per day
Low molecular weight antioxidant compound – Anoxe (2) (S. Jurasunas product)
18 gr per day
Fermented Chlorella
15 tablets per day
Pomegranate juice (target BCL2, BAX, VEGF, P21, NF.KB)

to drink during the day
1 – 2 – See the reference of the articles by Dr. Serge Jurasunas, either in English or German. 73
F. 62 years old – Advanced uterine cancer with neoplasic
infiltrative lesions - 2009
Before
Treatment - chemotherapy
Angiogenic therapy
L.C.E
Biological Response Modifier
Biobran MGn3
Modulate oxidative stress and apoptosis
(NF.KB – Cox 2
Curcuma – Fermented Chlorella
Low molecular weight
Antioxidant compound Anoxe
After
Anticancer diet
After 4 months complete therapeutic response

with a total regression of the large uterine
tumor of 6 cm and complete regression of all
the adenopathies (2cm) from the wall of the
rectum and vaginal canal.
Good quality of life during chemotherapy

74
Actived P53 gene expression – High protein level
Effects of Anoxe – A low molecular antioxidant
compound on a resistant tumor to radiation therapy
M. 42 years – lymphoma treated with

chemotherapy.
Recurrency with a large tumor

localized in the bronchial area
resistant to new radiation therapy.
Before treatment
40 days using the low molecular

antioxidant compound Anoxe
(together with radiation) at 18g per
day + LCE (14 ml per day).
Shown a significative reduction of the
tumor size.
After treatment 75
Dr. Serge Jurasunas
Patient female –50 years old with a metastasic recurrency from breast
cancer (2000) in 2004 with multiple lesions dissiminated in the skeleton.
Strong pains and after one year of a new protocol of chemotherapy a new
bone scan (May 2005) showed more lesions disseminated in comparaition
of 2004. Poor response and evolution of the disease.
The complete case fully illustrated is available in English and French.
X ray of the spine
After 3 months of
combined therapy +
chemotherapy.
Total elimination of the

very large lesions in the
illium and spine.
76
Jurasunas Serge, ND
Old Case
Breast cancer survivor
1994 – Age 36 years. June 2008 – 50 years

Bad prognostic of 18 ganglions Participating as witness support in
with metastasis – Liver and bone our meeting group of psychological
metastasis support of cancer patients
77
Emerging paradigm shifts
■ individualized treatment based on tumor and patient profiling.
■ New tumor markers for early detection, monitoring of treatment,

predictive value.
■ Assess cancer cells resistance from overexpressed anti-apoptotic

genes.
■ Evaluate the protumoral activity over the antitumoral (vice versa).
■ Evaluate or anticipated risk of recurrency.
■ Combination treatment introducing Dietary agents and other safe

supplementations that may target the different Hallmark of cancer.
78
Strategies for Efficient Cancer Therapy
Dietary agents and natural compounds to target P53,
BCL-2, BAX, Survivin, P21-like-proteins and increase
chemotherapy/radiation effectiveness
Dietary agents:
Resveratrol, Pomegranate, Curcumin, Indole-3-carbinol, Ellagic acid, Quercitin,
Green-tea, etc.
Food – Anticancer diet:

Cruciferous vegetables – Apigenin, Isithiocynates, Sulforaphane
Natural compounds:
Oligopeptide (short chain of amino-acids extract from fish)
Fermented chlorella - Low molecular antioxidants compounds (Anoxe)
Enzyme yeast cells preparation– Omega 3 – Curcumin – Resveratrol –
Biobran MGn3
Bharat B., Aggarwel, Shishir Shisbadia – Molecular targets of dietary agents for prevention
and therapy of cancer – Cytokine research lab – Dept Exp therapeutics – Univ. Texas M.D.
Anderson Cancer Center – Texas (USA). 79
Chemical structure of dietary agents
Dietary agents with

anticanter properties
Chemical structures of dietary

compounds
80
81
Chemotherapy + Radiation + Dietary agents
Why Dietary agents?
Cisplatin Well tolerated

Doxorubicin by the body
Docetaxel, etc. Induce
apoptosis Selective
Chemotherapy Dietary agents
Quercitin
Radiation Resveratrol
Curcumin
Non selective Pomegranate
Genistan Non-toxicity
Adverse toxic effects
Inhibit NF.KB
Apoptosis
Synergy to increase
effectiveness of Decrease adverse
anticancer agents toxic effects
82
Chendil D. et al – Oncogene 2004: 26: 1599 - 607
Curcumin potentiates the effect of paclitaxel by suppressing
the metastasis of the human breast cancer to the lung in
mouse xenogragraft model
83
Articles about how to target mutant P53 and target
molecular markers by
Serge Jurasunas
1 – How to target mutant P53 in a case of multiple cancer recurrence

– Townsend Letter August/Sept 2010-325-26
2 – Gezieltes Vorgehen gegen mutiertes P53 bei multiplen Krebsrezidiven

- Co-Med nº 5/2010
3 – Breast cancer: Prevention, Detection, Targeting Molecular Markers

- Townsend Letter August/Sept 2011
4 – Brustkrebsdiagnostik und prävention – Prävention durch P53

tumorsuppressorgen und Molecular Marker
- Co-Med Mai/2012
84
Modulation of Apoptosis by active compounds for cancer therapy
Organ Site Molecular target

Curcumin Lung, pancreas, colon, prostate, P53 – P21 – Survivin – NF.KB –
stomach, breast. Cox2 – VEGF – BAX – BCL-2
Increase chemotherapy sensitivity-

synergetic
Gemcitabline, oxilaplatin,
paclitaxel
5 Fu
Resveratrol Ovarian, breast, prostate, liver, P53 – BAX – Caspases activator –

lung, uterine. P21 – Survivin – BCL-2 – Cyclin 1 –
Cisplatin, doxorubicin, platinum Cox2
compounds, 5 Fu, paclitaxel G1-G2 – S-phase arrest
Apigenin Pancreatic cancer P53 – P21 – BCL-2 – BAX –

5Fu, Paclitaxel Caspases – C-Myc – VEGF –
induce G 2/ M – Cell cycle arrest
Pomegranate Prostate, breast, lung, colon, oral, BCL-2 – BAX – VEGF – Cox2 – P21
leukemia – BAK – NF.KB
Breast cancer, prostate, pancreatic BCL2 – BAX – NF.KB

Gemcitabine – cisplatin - paclitaxel Activate caspases
Indole-3 - Carbinol
85

Ellagic acid Neuroblastoma, pancreas, breast, P53 – P21 – Cyclin 1 – Cox 2 –
prostate, colon, intestine, bladder, NF.KB
oral, liver. Increase caspase 3
Cisplatin, vinorelbine
Quercitin Colon cancer, breast. BAX – BCL-2 – Caspase - Cyclin

D1 – Gadd 45
5Fu, Cisplatin
Doxorubicin
BCL-2 – BAX - Survivin – Cyclin D1
Capsaicin
NF-KB – CDK1 – CDK2
Increase Cyt C. release Caspase 3
Catechin 5 Fu, Gemcitabine – mitocycin
P53 – BCL-2 – P21

VEGF – Cyclin D1
86

Green tea Lung, oral, stomach, liver,
pancreas, colon, bladder, prostate,
Polyphenols and EGCG breast. P53 – P21 – BAX – BCL-2 – NF.KB
– Cyclin 1 - Cox2 – VEGF _
Tamoxifen – cisplatin – increase Xiap
adriamycin, dacarbazine
Isithiocyanates (ITC’s)
Prostate, breast
Docetaxel BCL-2 – Survivin – NF.KB – Cyclin
Adriamycin 1 – P53 – caspase activation
Sulforaphane Cisplatin
(Cruciferous vegetables)
Licopene Prostate, lung, breast, gastric,

liver, pancreas colorectal
Adriamycin Cyclin D1 - BCL-2 – BCL-4 AKT –
Cisplatin NF.KB – MMP9
87
Thank you for your attention
For more information, conference, clinical cases

results consult:
E-mail: info@sergejurasunas.com
88

Conf Munique 2012 - Professor Serge Jurasunas

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Target Therapies

Speaker: Serge Jurasunas M.D. (Hom) N.D.

Topic: How to treat cancer by targeting P53, BCL-2, Survivin:

Integrative Medicine – Complementary Oncology

Our work focus 3 directions:

● Cancer prevention (earlier diagnosis)

● Prevention of disease recurrence

Complementary Test: Live blood analysis, thermography, electromagnetic

Treatment: immunomodulation, angiogenic therapy, antioxidant therapy, stem

● Cancer a matter of life or death.

● Clinical application of molecular markers and their proteins with cancer

● Dietary foods and compounds to target gene expression.

● Cancer cell survival is totally

● The balance between pro-

■ For a population of cells to exhibit

P53 is the “Guardian of the genome” which instructs

Thus, P53 guards against changes to cells that might lead to

P53 mutation or inactivatition appears necessary to develop

P53 is mutated in about half of all tumors.

P53 gene is one of the most

Gain of Function (G.O.F)

Mutagenesis Dominant negative effects

Pro-apoptotic signals via BH3 only

■ One half of the invasive ductal

■ Increasing evidence that

■ Distant metastases express

NFKB Cytosol Release NFKB translocates to the nucleus and

■ Associated with increasing cancer cell resistance to chemotherapy or

■ Diagnostic Anticancer Strategy

■ Re-establish chemo sensitivity

Several basic elements are involved in the malignant process

■ Elevated NF.KB activity

■ Activated Pi3 – Kinase pathway

90% of human cancer deaths are caused by metastases

Reactivating Mutant P53 using small molecules

Gain of function Normal function Tumor

■ BCL-2 enhances angiogenesis

A layer of pink normal cells is shown

Pink – normal cells

Tumor cells are not undergoing apoptosis 24

A key player in cell death induced by anticancer drugs

■ Bax gene expression is inactive in approximately one-third of invasive

■ Enhanced expression of Bax proteins is associated with good response to

- Activates the proapoptotic

- induce the formation of the

- APAF1 – cytochrome C complex

Cells that overexpress BAX enhance cell death.

Cells that overexpress BCL-2 reduce the suceptibility to apoptosis.

Because BAX proteins antagonize BCL-2’s anti-apoptotic function, it

Scopa, Chriscula, Vagionas, Constantine, Kardimaki, Dimitris, Kourelis, Athanasias C. –

The effects of zinc on Bax and Bcl-2 levels of

Survivin is an inhibitor of apoptotic proteins (IAP’s) inhibit Caspase 3,

Survivin expression is absent normal tissue however overexpressed

WT P53 may transcriptionaly repress Survivin gene expression P53

Extracellular Survivin released is promptly captured by neighboring

■ Nuclear cytoplasmic and mitochondrial pools of Survivin (Fortugno

■ Nuclear localization is linked with the capacity of Survivin to

■ Extracellular Survivin released is promptly

■ T-cells mount a vigourous cytotoxic response to

Anderson, MH et al (2001) – Identification of a cytotoxic T-

■ Survivin is strongly up-regulated in angiogenically stimulated

■ increased Survivin expression protected E.C. from Apoptosis during the

■ Targeting Survivin in E.C. may be a potential strategy to inhibit – tumor