A homeopathic bit of breast cancer science, or Who knew
Cytotoxic effects of ultra-diluted remedies
alcohol was so toxic Respectful Insolence.mht
A "homeopathic" bit of breast cancer "science,"
or: Who knew alcohol was so toxic?
Category: Alternative medicine Medicine
Quackery
Posted on: March 4, 2010 9:00 AM, by Orac
Homeopaths are irritating.
They're irritating for a number of reasons. One is
their magical thinking, and, make no mistake, their
thinking is nothing but pure magic, sympathetic
magic to be precise. That's all that the principle of
"like cures like" really is at its heart. Normally, that
principle states that "like produces like," but
homeopathy reverses that principle by saying that,
while like produces like at normal concentrations,
like reverses like at high dilutions. Somehow the
magical process of shaking the remedy very hard
between each dilution step (called "succussion")
imbues it with the property of curing what it would
normally cause. Of course, scientists know that the
whole "shake and dilute" process is bunk, given that
a typical 30C homeopathic dilution dilutes a
substance to the point where it is incredibly unlikely
that even a single molecule remains.
This brings us to the second magical principle upon
which homeopathy rests: The law of contagion,
which states that things that have once been in
contact with each other continue to act on each
other at a distance after the physical contact has
been severed. The perfect embodiment of this is the
claimed "memory of water," which is how
homeopaths get around the inconvient fact that most
of their remedies contain not a single molecule of
the original remedy. Somehow the water
"remembers" the contact with the remedy and
magically forgets the contact it's had with all the
other things that water has been contact with, like
poo, pee, chemicals, foodstuffs, and everything else.
It's not as if waterfalls and various other natural
processes don't "succuss" the water quite nicely.
What makes homeopaths particularly irritating is
that they believe so highly in their magic that they
all too often abuse science to try to "prove" that it
works. Lately, homeopaths have been throwing a
new study in my face as "evidence" that their woo
works:
Frenkel M, Mishra BM, Sen S, Yang P, Pawlus A,
Vence L, Leblanc A, Cohen L, Banerji P, Banerji P.
(2010).
on breast cancer cells. Int J Oncol. 2010
Feb;36(2):395-403.
Let's looks at the abstract:
The use of ultra-diluted natural products in the
management of disease and treatment of cancer has
generated a lot of interest and controversy. We
conducted an in vitro study to determine if products
prescribed by a clinic in India have any effect on
breast cancer cell lines. We studied four ultra-
diluted remedies (Carcinosin, Phytolacca, Conium
and Thuja) against two human breast
adenocarcinoma cell lines (MCF-7 and MDA-MB-
231) and a cell line derived from immortalized
normal human mammary epithelial cells (HMLE).
The remedies exerted preferential cytotoxic effects
against the two breast cancer cell lines, causing cell
cycle delay/arrest and apoptosis. These effects were
accompanied by altered expression of the cell cycle
regulatory proteins, including downregulation of
phosphorylated Rb and upregulation of the CDK
inhibitor p27, which were likely responsible for the
cell cycle delay/arrest as well as induction of the
apoptotic cascade that manifested in the activation
of caspase 7 and cleavage of PARP in the treated
cells. The findings demonstrate biological activity
of these natural products when presented at ultra-
diluted doses. Further in-depth studies with
additional cell lines and animal models are
warranted to explore the clinical applicability of
these agents.
Basically, it's a test of homeopathic remedies
against breast cancer cell lines in a dish that claims
differential toxicity aginast breast cancer cell lines
compared to normal breast cells and claim that
homeopathic remedies did just as well as cytotoxic
chemotherapy. For three weeks, I have been
assiduously ignoring them because I thought that
Dr. Rachie's takedown of the study was excellent
(not to mention that neither my university library
nor those of several of my friends apparently carry a
digital subscription to the journal--the best that
anyone could send me was from a library loan,
which was FAXED, and the figures come out as not
that clear), but yesterday was the last straw. I don't
know what it was, but finally I had had enough. Dr.
Rachie did good, but I'm a breast cancer researcher,
and I have worked with the cell lines used in the
paper. It's time for a heapin' helpin' of not-so-
Respectful Insolence. It's also time for me to make a
note never to submit a paper to the International
Journal of Oncology, whose editorial and peer
review standards are clearly lacking.
Dr. Rachie hit upon one huge flaw in the study, and
I can't help but mention it as well. There isn't a
single mention of statistics to show that the
differences described are significant. In fact, there is
a rather disturbing lack of proper quantification of
results throughout the paper, in particular with
image-driven data, such as assays for DNA
breakage by FISH. For these analyses, all the
authors show are pictures of "representative" cells,
but they didn't bother to analyze large numbers of
cells to see if the qualitative results that show up on
the panels they chose to print are real, if they hold
up to statistical analysis. It's very easy to be fooled,
even unintentionally, if you don't look at large
numbers of cells. The same is true of the flow
cytometry data, as Dr. Rachie also points out.
Let's step back a minute, now. This study is based
on homeopathic remedies of the P. Banerji
Homeopathic Research Foundation (homeopatic
research: an oxymoron if I've ever heard one!),
which claims to be able to cure a lot of cancers that
"conventional therapy" can't. Indeed, they even
submitted a "best case" series to the NIH in which,
out of 941 patients with breast cancer, the PBHRF
claimed that in 19% of patients, tumors regressed
and that 21% of tumors were stable. One thing I
noted is that the reference cited for this claim is not
a peer-reviewed article. It's merely cited as:
Banerji, P. P. Banerji Homeopathic Research
Foundation (PBHRF) in Kolkata, India, 2008.
Gee, I wish I could cite claims like that in my
papers!
Of course, the claim above is meaningless without
knowing a lot more about the patients. For instance,
as I've pointed out before, as many as 20% of
mammographically detected breast cancers might
spontaneously regress. I've also pointed out that the
natural history of breast cancer is highly variable,
from relatively rapid growth and decline to a slow
and indolent course. Indeed, a subset of women
with untreated breast cancer can survive as long as
10 years or more, and this data comes from before
the age of mammography, which means that these
were at the very least palpable tumors. Indeed, most
tumors detected at the end of the 19th century and
the beginning of the 20th century (which is when
this data was collected) were stage II and III. These
days, the vast majority of breast cancers are
detected by mammography and are stages 0
(noninvasive--or preinvasive--ductal carcinoma in
situ) or stage I. Without stage-specific survival data,
with stages documented by pathology, imaging
studies, and clinical data, a claim like the one made
above is uninterpretable and, in essence,
meaningless.
The remedies, however, are Carcinosin, 30C;
Conium maculatum, 3C; Phytolacca decandra,
200C and Thuja occidentalis, 30C. At this point I
can't help but note a couple of things. First, a 3C
dilution of conium muculatum is only a 106 or one
to one million dilution, which means that there
might well be something active left in this particular
remedy. The second point I'd like to make is just to
express awe at a 200C homeopathic dilution, which
would represent a dilution of 10400, or 10367 or so
(what's an order of magnitude between friends?)
greater than Avagadro's number. This is a mind-
boggling number, not to mention a mind-bogglingly
tedious process.
There's one other aspect to consider. The diluent
was not water. Rather, it was 87% "extra neutral
alcohol," and this alcohol was used as the control.
So far, fair enough. At least they used a solvent-only
control, which is more than I can say for a lot of
homeopathic "researchers." They then treated the
cells with different "doses" (although how one can
speak of a dose of a nonexistent remedy, I don't
know) of the remedies or solvent. The cells treated
included MCF-7 cells (which are estrogen receptor-
positive, or ER(+), cells and a very commonly used
cell line in breast cancer research; I use them
myself) and MDA-MB-231 cells (an ER(-),
progesterone receptor-negative, HER2/neu-negative
cell line, which I've also used in cell culture and
xenograft experiments). The former is relatively
slow-growing and somewhat tricky to grow in
xenografts, although easy to grow in cell culture,
and the latter is a model of a particularly nasty form
of breast cancer known as "triple negative" breast
cancer.. Control cells are an immortalized human
mammary epithelial cell line, HMLE, which are
supposed to represent "normal" mammary epithelial
cells.
I think the entire problem with the paper can be
summed up with this figure, which shows the effect
of these remedies on cell viability:
Notice how each remedy and the controls are
graphed in little bar graphs for each cell type. This
makes it very difficult to compare directly effects of
each remedy at each dose compared to the solvent
control. Really. Click on the picture and see if you
can easily tell whether the differences shown look
significant. You can't. Moreover, no statistics are
provided to tell you whether the differences might
be significant. Worse, however, look at the solvent-
only control groups. Do you see what I see? The
solvent is killing the cells, up to 50% of them! Not
only that, but the normal control cells appear to be
more resistant to the effect of the solvent, with very
little effect seen. In fact, by the lack of a clear dose-
response curve for the solvent-only control, with the
measurement fluctuating around a point slightly
lower than that seen with no solvent added, you can
get an idea of the variability of this assay from well
to well, which is quite a bit. The cynic in me thinks
that the data were graphed this way intentionally, to
obscure just how weak it is, but the angel in me will
attribute it to gross incompetence.
If I had reviewed this paper, I would have insisted
for the figure above that all the data for each cell
line be graphed on a single graph, with a dose-
response curve clearly indicated for each and error
bars, along with proper statistical analysis for a
dose-response effect showing that there were
significant differences between the effect of the
homeopathic remedies at each dose compared to the
solvent control. All that the above figure shows is
that adding 10 l/ml of 87% alcohol has a
significant effect on cell viability for the cancer cell
lines. They have not shown in any way that I can
tell that any of their remedies have a significant
effect on cell viability compared to the alcohol
solvent alone. As I've said time and time again,
anyone can kill cells in a dish with alcohol or bleach
or any number of other nonspecific solvents. That
does not mean that this study shows that these
homeopathic remedies are as "effective as
paclitaxel" (a commonly used chemotherapeutic
agent in breast cancer) against these cells. All it
proves is that the solvent at the doses used can
nonspecifically kill cells and that perhaps the
normal cells are more resistant to the alcohol.
Speaking of the solvent, there's something very
fishy in this passage from the paper:
All four remedies had very similar HPLC
chromatograms to each other, with only trace
amounts of limited number of peaks. They were not
significantly distinct from the solvent and they
lacked the distinct peak seen in the solvent.
The chromatograms are not shown, but note the last
part: They lacked the distinct peak seen in the
solvent. What on earth does this mean? One guess is
that the solvent peak was wider and less distinct in
the homeopathic remedies, but another is that it was
gone. Which is it? Who knows? As I said before,
the chromatograms weren't shown. In any case, if
these remedies are truly homeopathic, then why are
the solvent peaks different?
I seriously have to wonder about this solvent. The
maximum dose used (10 l/ml) is in essence a
1:100 dilution of alcohol, making the alcohol
content in the media less than 1%. The lowest dose
(1.25 l/ml), is close to a 1:1000 dilution. Alcohol
alone should not be that toxic to these cells. At least,
in our hands it isn't. We've used drugs diluted in
alcohol at similar concentrations to the lowest dose
of solvent used in this study (1.25 l/ml), and we
don't see 35% cell kill in MDA-MB-231 or MCF-7
cells due to solvent effects alone. Something odd is
going on here.
A journalist acquaintance of mine once asked me
for examples of massive failures in the peer review
process. I can't think of a better one than this paper.
It tests a remedy so highly implausible as to be
safely considered, for all practical intents and
purposes, impossible barring some truly
extraordinary evidence coming to light, evidence
sufficient to overthrow long-established science in
multiple disciplines. Its statistical analysis is
nonexistent, and its quantification dodgy in many
places. All of this means that its conclusions do not
flow from its data and are not supported by its data.
The only conclusion that is supported by the data is
that the solvent in which the homeopathic remedies
had been diluted is toxic to MCF-7 and MDA-MB-
231 cells.
Finally, I can accept that perhaps a 3C homeopathic
dilution might have an effect on cells. There could
be an actual drug remaining there. However, 30C
and 200C homeopathic dilutions leave nothing
behind, and there is nothing in this paper to show
that there is an effect above and beyond solvent
effects from either of these remedies. Come to think
of it, there's nothing in this paper to show that the
3C homeopathic dilution really has any effect above
and beyond solvent toxicity effects. Put it all
together, and this is a EPIC FAIL on the part of the
peer reviewers and the editors.
You know what's more irritating than homeopaths?
Peer reviewers and editors who let dreck like this
see print, giving homeopaths something to irritate
me with.