Journal of Pharmacy and Nutrition Sciences, 2015, 5, 000-000 1
ISSN: 2223-3806 / E-ISSN: 1927-5951/15 © 2015 Lifescience G lobal
Can Plants’ Ability for DNA Repair and Stress Defense be Used
against Patients’ Circulating Tumor Cells?
C.D. Volko and U.D. Rohr*
Endobal Medical Research Company, 1103 E. Tropicana Ave. #3006, Las Vegas, Nevada 89119, USA
Abstract: Introduction: It w as suggested that specific plants may reduce canc er's r esistance to c hemotherapy .
Resistance inhib its apo ptosis, as well as other fundamental an ti-cancer protective mechanisms. Soy bean has been
found to reduce cellular stress and re pair DNA dama ge caused by drought or parasit es, and can tr ansfer t his def ense
mechanism to other plant species as well. The aim of this study is therefore to conduct a systematic comparison of the
effect of s oy bean formulation (FSWW08) on gene expression i n in vitro human breast cancer cell line, and in in vivo in
blood circulating t umor ce lls (CTC), after oral consumptio n of FSWW08 by patients suffering from breast-, ovar ian-, and
Method: In vitro gene expressions studies were conducted wit h the human breast cancer c ell line BT-474 that was
exposed to doxorubicin or FSWW08, either alone or in combination. Ovarian-, prostate-, and breast cance r patients
received FSWW08 for 30 days. CTC were extracted from their blood according to an established protocol. Gene
expression evaluations were conducted before and after treatment.
Results: In vitro, the multi-drug resistance (MDR) protein was reduced by FSWW08, but was increased by doxorubicin.
The combination of F SWW08 and doxorubicin, however, showed a protective effect against the increase of MDR in
physiologic conce ntrations, increased, however, also in high experimental co ncentrat ions of bo th agents. The expr essio n
of several cancer-related protective genes, such as tumor suppressor factors p21, p38 and p 53, was improved by
FSWW08 in vitro and in vivo, which helped cell differentiation and new tissue formation. Additionally, the BAX/Bcl2 ratio
was improv ed, in vit ro, as well as gene expression of estrogen recept or beta, NF-B, MAP kinase, c-JUN, and matrix
metalloproteinase 9, together with an increase of VEGF expression in vivo in CTC.
Conclusion: It was demonstrated that FSWW08 improved the gene functions related to DNA re pair and s tress in hu man
blood CTC and resistance marker, in vitro, when applied in combination with doxorubicin. As such, larger in vitro and in
vivo clinical stu dies that investigate single botanical compounds from other pl ants, ar e warranted.
Keywords: Tp53, Tp21, Bax/Bcl2, MAP kinase, VEGF, CTC, circulating tumor cell, fermented soy, MDR protein,
estrogen receptor beta, NF-B, RT-PCR technique, human breast cancer cell line BT-474, ovarian cancer, breast
cancer, prostate cancer.
During World War II, the US government developed
protective measures against cancer . This served as
the model for a long series of similar agents (alkylating
agents) killing rapidly growing cancer cells by
damaging their DNA - the beginning of chemotherapy
According to experts in the field two problems have
to be solved to improve chemotherapy: Resistance and
side effects [2-6]. Several new approaches are now
being studied, including new drugs, new combinations
of drugs, new delivery techniques, novel approaches
that target drugs more specifically at the cancer cells,
first, drugs to reduce side effects and second, agents
that overcome multi-drug resistance [2-6]. Several
reviewers suggested that plant based formulations may
inhibit resistance against cancer therapy, including
Isoflavones, curcuma, vitamin A, and some extracted
ingredients from Chinese herbs .
*Address correspondenc e to this author at the 1101 E. Tropicana Ave. #3006 ,
Las Vegas, Nevada 89119, USA; Tel: 1 202 321 6621;
A new concept suggests that cancer develops from
a small group of cells similar to stem cells, which
means that they are not fully differentiated, show
impaired hormone receptor expression (Figure 1) and
are both the origin of cancer (Figure 2) and responsible
for metastasis through migration (Figure 3) . The
cancer stem cell theory has profound implications for
cancer chemotherapy because it explains why it is
effective in more differentiated cells but ineffective in
cancer stem cells . The spread of cancer is
facilitated via migration of the blood (circulating tumor
cells or CTC) or the lymph system to other organs
(Figure 3). It was suggested that plant based natural
agents may be particularly effective managing CTC,
which was shown in vitro . Several chemotherapeutic
agents, which do stop fast growing tumors, have the
opposite effect on cancer stem cells, causing them to
divide too rapidly , as well as increasing stress
related oxygen species (ROS), causing resistance to
the therapy, whereas some plant ingredients are able
to reduce ROS and reduce stress resistance to cancer
Scientists explain human cancers which sustained
proliferation, escape from apoptosis, and genomic
2 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
instability by mutations of the TP53, ATM (ataxia-
telangiectasia mutated (ATM) gene, and MDM2
(Mouse double minute 2 homolog) genes . It is
defined that DNA replication stress generates genomic
instability and causes suppressed apoptosis [10-21].
Figure 1: Hormone receptor distribution in stem cells, cancer
stem cells, and normal fully differentiated Schwann cells,
taken from .
Recently, researchers have suggested that dietary
compounds such as curcumin, sulforaphana, soy
Isoflavones, epigallocatechin-3-gallate, resveratrol,
lycopene, piperine and vitamin D3 can directly or
indirectly affect cancer stem cell self-renewal pathways
, as well as reduce unfavorable cellular stress
caused by chemotherapy and making chemotherapy
more effective [23, 24]. Plants have evolved to live in
environments where they are often exposed to different
stress factors in combination and are subject to high
levels of DNA damage resulting from the plant’s
obligatory dependence on environmental stresses like
solar UV radiation, attacks of yeasts to digest plants,
drought, chilling injury, and other air and soil pollutants
including heavy metals and metabolic by-products from
endogenous processes . Therefore, to survive
under frequent and extreme environmental stress
conditions, plant cells have evolved with highly efﬁcient
and wide-ranging mechanisms for the detection and
repair of DNA damage to eliminate the chances of
permanent genetic alterations and to maintain genome
stability for faithful transfer of genetic information over
Classically plant based anticancer medicine is toxic,
often inhibits mitosis, follows the first generation
anticancer drugs made from mustard gas and has
severe side effects. The here outlined mechanism of
DNA repair differs completely from this strategy: it is a
non-toxic mechanism, since plants under
environmental stress attack are already severely
immunologically compromised and need no further
reduction of defense mechanisms  The DNA repair
under stress is a mechanism to strengthen impaired
plants. Yeasts and plants share genes for cell repair
and DNA protection with mammals like MAPkinase and
heat shock protein (Figure 4) . MAPkinase
cascades are crucial in eukaryotes for transducing the
perception of environmental stimuli into internal
signaling pathways . Plants have a particularly
Figure 2: Schematic depiction of invasive breast cancer. Primary breast cancer consists of two types of cancer cells. Cancer
stem cells have only low estrogen receptor expression, whereas primary cancer consists of cancer cells expressing 62%
estrogen receptor. Chemo-, radio-, or adjuvant therapy cannot reduce cancer stem cells, because they have virtually no
estrogen receptor expression. Therefore, cancer recurrence is difficult to predict. (See ref ).
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 3
large number of MAPkinase components, allowing for
the control over a wide range of stress response
pathways . Interestingly, in Arabidopsis, a unique
plant-speciﬁc transcription factor Suppressor of
Gamma Response 1 (SOG1) acts as the main
regulator in DNA damage repair and performs
analogous functions to mammalian tp53, which is
involved in a majority of the plant’s response to DNA
damage, such as transcriptional response, activation of
cell cycle checkpoint and programmed death of stem
cells . It is a tremendously important finding that in
plants a factor similar to tumor suppressor factor tp53
exists, which is a hallmark gene factor for cell
differentiation in mammals, since in more than 50% of
all tumors tp53 is missing . Transcription analyses
in Arabidopsis plants and many others have revealed
that a fairly high proportion of the genes are associated
with DNA damage repair [34, 35].
Transcription factors are controlled by plant
hormones and small proteins (Figure 4): The
expression of soybean miRNAs was investigated in
response to water deﬁcit and infection with soybean
rust fungus, where all miRNAs were differentially
regulated by each stress, usually in an opposite
direction . It has been shown that soybeans are
able to affect stress response in other species too, like
tomato plants as well as tobacco plants . There are
numerous studies showing that soy can alter the
genome in human cancer cells . This paper
therefore systematically investigates the effect of
fermented soy bean formulation on cancer cells in vitro,
Figure 3: Tumor spread is facilitated via cancer stem cells, which have very low estrogen receptor expression and might not
respond to chemo-, radio-, or adjuvant therapy.
Figure 4: Key events in the signal transduction pathway
activated in response to combined biotic and abiotic stresses
(taken from ref. ).
4 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
as was shown by others also, in vivo in CTC extracted
from blood from cancer patients, which has not been
shown before. We detected and reported earlier that
the effectiveness of chemotherapy was increased by
fermented soy formulation, side effects were reduced,
and survival increased [39, 40]. This study investigates
in vitro whether a combination of the classic cytotoxic
compound doxorubicin may be combined with
fermented soy bean and counterbalance some of the
side effects of classical chemotherapy on the genetic
level. For the first time ever stress DNA repair genes
are investigated in blood circulating tumor cells (CTC)
in vivo altered by a processed plant.
MATERIALS AND METHODS
The study protocol was reviewed by a local
institutional review board (IRB) to protect the rights of
patients, safety and well-being of humans involved in a
clinical trial by reviewing all aspects of the trial and
approving its startup and was published previously [24,
39, 40]. All patients had to sign an Informed Consent
Form after the physician explained the study and he
read the informed consent form where the study goal
was outlined in a written form, in which the study was
outlined and participation was voluntarily and it was
explained individually to all patients that they could
leave the study at any time even without explanation.
No patient received a financial compensation for
participating in the study, besides received the
medication for free. The study fulfilled the Convention
of Helsinki in its current form. The data were stored at
the Hospital and the data were processed anonymous
to protect the identity of the patient.
b. In Vitro Gene Expression Investigation
All in vitro procedures determining the gene
expression determination have been described before
[24, 39, 40, 44], however a more thorough investigation
and discussion about in vi tro and in vivo comparison
and DNA repair is missing. Investigations of the human
breast cancer cell line BT-474 were conducted, which
were purchased from the German National Resource
Centre for Biological Material (DSMZ, Leibniz Institute
DSMZ-German Collection of Microorganisms and Cell
Cultures, Inhoffenstraße 7B, 38124 Braunschweig,
GERMANY, EU). It is a human breast tumor-cell line
and was established from a solid invasive ductal
carcinoma of the breast (DSMZ No.: 64.
Doxorubicin hydrochloride was solved as 10 mg in 5
ml volume aqueous solution and purchased from
HEXAL AG, 83607 Holzkirchen, Germany, EU.
FSWW08 (Haelan 951, Organic NON-GMO Soy), a
fermented Soy Bean Beverage made in China, is
distributed by Haelan Products INC., 18568 142nd Ave.
N.E. Bldg.F. Woodinville, WA 98072 U.S.A.,
www.haelan951.com. FSWW08 was given as a
donation. FSWW08 contains 250 mg soy Isoflavones
A RPMI Medium with FBS was employed according
to the recommended condition of DSMZ Cell Culture
Data. For every test 1x105 BT-474 tumor cells were
used. Growth was induced on a 24 Well-plate
(CELLSTAR) purchased by Greiner bio-one. After
adding the drug substances, the cells were incubated
over a period of 72 h at a temperature of 37° Celsius in
Processing of Harvested Cells
Cells were processed according to recommended
procedures by Quiagen with a QIAmp RNA Blood Mini
Kit to obtain total-RNA. This was obtained by
cryopreservation and Freeze-Drying Protocols of the
cell pellet with Quiagen RLT-buffer. Further isolation of
RNA was performed by QIA shredder column and QIA
spin column. The RNA containing elute was evaluated
by reverse transcription. The reaction was conducted
with the help of random hexamers and M-MLV
transcriptase, so that RNA was transformed to cDNA.
Quantitative Real-Time RT-PCR was employed to
measure mRNA gene expression. Gene expression
measurements were conducted with Sequence
Detector 7700 by Applied Biosystems (ABI) Life
Technologies, 3175 Staley Rd, Grand Island, NY
14072, USA. A 5´-Nuclease Assay was employed,
which was developed by ABI. The method is based on
measurement of quantitative determination of cDNA as
an equivalent amount of mRNA. For quantitative
purposes every gene expression determination was
done with a control cell line, which contained mRNA as
a standard. The dilution of the control cell line was
done in ratio of 1:10.
The measurement was done in a relation as a
fraction to an amount of cDNA 2x106. The value
obtained in the Real-Time RT-PCR is the cell
equivalent (CEQ). All measurements were done in
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 5
The measured gene expression determination was
related to GAPDH and normalized to allow comparison
among each other. As an additional method all
obtained values were normalized to 1 as an untreated
tumor, so that a relative gene expression is expressed
in all figures.
Dosing of Test Substances
Doxorubicin is used in cancer mono therapy usually
in a dose of 50-80mg/m body surface. The average
body surface of an adult is usually 2 m. The injection
solution is composed of 2μg Doxorubicin/μl. The
amount of blood of an adult person is 5-7 Liter on the
average. The dose of FSWW08 is one bottle containing
236 ml per day, half a dose 118 ml twice a day.
Substances were tested alone and in combination and
compared with an untreated cell line. Gene expression
of parameters (BCL2, BAX, TopoII, MDR1, ERBB2 and
others, Table 2) in different concentrations were
measured. The questions to answer are:
a) What are the effects of Doxorubicin monotherapy
on the expression of genes related to the
mamma carcinoma cell line BT-474?
b) What are the effects of the combination of
Doxorubicin and FSWW08 on the gene
expression in the cell line BT-474?
c) Is it possible to obtain similar results when
combining Doxorubicin with FSWW08 as in
using Doxorubicin alone on BT-474?
d) In vivo gene expression of tumor cells circulating
in blood extracted from patients
A pilot study was conducted with 5 ovarian cancer
patients, 5 prostate cancer patients, and 7 breast
cancer patients (pT1 to pT4, pN0 to PN2, Mo).
Two samples of 5 ml ethylenediamine tetraacetic
acid blood were collected for isolation of CTCs before
the application of therapeutic substances with an S-
Monovette® (Sarstedt AG & Co, Nürmbrecht,
Germany) and were stored at 4°C until further
examination. The samples were processed
immediately or not later than 4 hours after blood
withdrawal. An additional serum sample was collected
to determine serum tumor markers.
Extracting Cancer Tumor Cells from Blood
The methods are described in .
Method of Determination of Gene Expression
The method is described above as in vitro and was
used with the same laboratory protocols and
equipment. The lower detection limit of this assay is
based on spiking experiments two cells per 5 ml .
Examination of blood samples of healthy donors (n =
20) yielded a specificity of 95%, which corroborates
values in the literature . We detected in all ovarian
(100%), all prostate cancer (100%) as well as in 5 from
7 women suffering from breast cancer CTC (71%),
which is higher than reported in the literature .
c. Gene Expression in CTC in Cancer Patients
Extracted from Blood
We relate gene expression results to two clinical
trials, where we investigated a) in a pilot trial over a 4
year period survival  and well-being of prostate-,
breast- and ovarian cancer, and b) in a multicenter
placebo controlled trial in 268 patients . We
compared FSWW08 with a casein solution. Besides
cachexia, white blood counts and other blood
parameters were determined. Patient characteristics
can be found in the literature [39, 40].
A t-test compares the means of two groups. For
example, compare whether relative gene expression
differs between a control and treated group. The t test
compares one variable between two groups. The t-
tests (and related nonparametric tests) compare
exactly two groups. Software was used of the 2014
version of GraphPad Software, Inc., 7825 Fay Avenue,
Suite 230, La Jolla, CA 92037 USA,
The effects of Doxorubicin and FSWW08 on gene
expression on GAPDH of BT-474 cells were
investigated separately and compared to untreated BT-
474 cells (Table 1) (Figure 5). Both Doxorubicin and
FSWW08 resulted in reduction of the cell morphology
and gene expression of BT-474 (Table 1, Figure 5).
Doxorubicin: The expression of GAPDH was
decreased by 50% with low DOX concentration and by
more than 90% at higher DOX concentrations. This
implies a significant reduction of the number of
surviving tumor cells (Table 1, Figure 5). FSWW08:
FSWW08 resulted in a 5-fold decrease of GAPDH
gene expression compared to untreated tumor cells
and caused up to a 10-fold decrease. This means that
FSWW08 had a significant effect on the number of
surviving tumor cells. Both Doxorubicin and FSWW08
have to be considered cytotoxic (Table 1, Figure 5).
6 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
Table 1: Summary of in vitro and in vivo gene expression changes by FSWW08. In vitro investigation were conducted
with normalized numbers of cells from cell line BT-474, purchased from the German National Resource
Centre for Biological Material (DSMZ, Leibniz Institute DSMZ-German Collection of Microorganisms and Cell
Cultures, Inhoffenstraße 7B, 38124 Braunschweig, G
GENE Function Cell Line
GADPH GAPDH is a house-keeping gene. Gen eexpression is equal
in tumor cells and in healthy cells. It is a gene for
normalisation and quantification because it is expressed only
in living c ells. A reduction of GADPH means that cells have
BAX BAX is a pro-apoptotic mitochondrial membrane protein. It
inhibits Bcl2 an d accelerates the programmed cell d eath
(apoptosis). Re duced expressi on of BAX in re lation to Bcl2
correlates with non-response to 5-flu orouracil, epirubicin and
Up to 4 fold No change 8 to 60 fold
Bcl2 Bcl2 is coding for an anti-apoptotic mitochondrial membrane
protein. Bc l 2 is overexpressed in many tumors and
consequently causes resistance to apoptosis-inducing drugs
(e.g. intercalat ing agents, alkylating agents, plati num
No change at
No change No change No
ERBB2 Erbb2 (also HER2/neu) is a tyrosine kinase involved in cell
proliferation and differentiation. Tumors overexpressing Erb b2
can be treated with Herceptin (an Erbb2-anti body).
No change No change No
MDR1 Multidrug r esistance 1 (MDR 1) is a glycoprotein cap able to
transport anticancer drugs out of the cells. Tumor cells
overexpressing MDR1 are resistant to multi ple dru g types like
vinca-alkaloides, anthracyclines, taxanes or mitomycin C.
-44 % - 25%
Tp21 The cyclin- dependent kinase inhi bitor p21 (als o known as
p21WAF1/Cip1) promotes cell cycle arrest in response to many
stimuli. It is well positioned to function as both a sensor and
an effector of multiple anti-pr oliferative si gnals.
tp53 tp53 is a tumor suppressor gene and regulates the cell cycle.
Mutations in p53 are the most frequent genetic alterations in
human mali gnancies. Genetic alterations in p53 (mutati ons,
LOH, expression) are therefore used as molecular tumor
c-jun c-Jun is an immunity mark er and is involved in ma ny allerg ic
and inflammatory reactions
Estrogen receptor beta is related to lower prostate c ancer
however its meaning in breast and in ovarian cancer is
The classical estrogen receptor relat ed to breast c ancer
survival, against tamoxife n protects.
No change No change No change No
Topo IIa Topoisomerase II alpha is catalyzing controlled cu ts and
reconnection of DNA-double strands during DNA- replication.
Inhibitors of Topoisomerase II (anthrazyclines, mitoxantron,
etoposid) are provoking faulty action of Top o II, leaving
behind DNA-damages. If Topo II is under expressed in tu mor
cells, lesser DNA-damages occur and the therapy may fail.
On the ot her han d, over expression of Topo II sensitizes the
cells to Topo II inhibitors.
ND ND ND
VEGF Vascular endothe lial gr owth factor (VEGF) is up-regulated in
many tumors and is inducing angiogenesis by paracrine
stimulati on of endothelial cells. High expression lev els of
VEGF in tumor cells are associated with poor response to
hormone therapy with t amoxif en and c ombination
chemotherapy with FAC or CMF. New anticancer agents like
bevacizumab (Avastin) are targeted s pecifically against
± 0 % 37 fold
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 7
Figure 5: Effects of Doxorubicin and fermented soy (FSWW08) on the GADPH expression of cell line BT-474. GADPH gene
expression of untreated cells expressed as 1.
Figure 6: The role of c-myc, Bax and Bcl2 in apoptosis. c-myc: a regulator gene that codes for a transcription factor, a mutated
version of which is found in many cancers. NF-kB: a protein complex that controls the transcription of DNA, important in stress,
immunity and cancers.
Change in the gene expression of anti-apoptotic
BCL2 (Figure 6): Doxorubicin: The effect of
Doxorubicin on the expression of BCL2 was very
strong. With increasing concentration of DOX the BCL2
expression values dwindled down to less than 10% of
what was found in the untreated BT-474 tumor cells
8 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
(only reported). FSWW08: FSWW08 had at low
concentrations no effect on regulation of the anti-
apoptotic BCL2 expression values (Table 1).
BAX gene expression (Table 1, Figure 7).
Doxorubicin had a low effect on the pro-apoptotic BAX
expression (not shown here, only reported). Application
of FSWW08 on the cell line BT-474 led to an increase
of pro-apoptotic (Figure 6) BAX expression, and the
increase of BAX expression was even higher: almost
the 4-fold number was achieved compared to untreated
TOPO IIa gene expression with increasing
concentration of agents: Doxorubicin led to a 1.8-fold
increase of TOPO IIa gene expression, gene
expression diminished again at high concentrations
(Table 1). TOPO IIa gene expression displayed only
the 0.6-fold value of untreated BT-474 cells. FSWW08
does not cause increase of TOPO IIa. On the contrary,
it causes an up to 0.3-fold decrease (Table 1).
MDR1 (multidrug resistance) gene expression
(Table 1, Figure 8), a marker of tumor resistance. An
increase of Doxorubicin led to significant increase of
the expression of the MDR1 gene, up to 17-fold
increase in the gene expression of MDR1. The
expression in the low concentrations of FSWW08 alone
decreased by 40%, comparable to untreated tumor
Figure 7: The effect of Doxorubicin (D) and FSWW08 on BAX gene expression in breast tumor cell line BT474. BAX gene
expression of untreated cells expressed as 1.
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 9
Figure 8: The effect of doxorubicin and fermented soy on
MDR1 gene expression (multi-drug resistance) in breast
tumor cell line BT474. MDR1 is a cell membrane protein that
pumps substances out of cells. MDR1 gene expression of
untreated cells expressed as 1. Upper figure doxorubicin is
added. Lower Figure doxorubicin is added and FSWW08 in
increasing concentrations, denoted at the x-axis.
Combination of Doxorubicin and FSWW08
With the low concentration of 10 DOX, an
increasing concentration of FSWW08 also caused an
up regulation of the expression of MDR1. With 10
DOX/60 FSWW08 it was possible to achieve an
expression corresponding to a 60 FSWW08 mono
therapy. A high concentration of Doxorubicin 100 DOX
combined with 10 HAL had an effect comparable to
Doxorubicin mono treatment. Increasing the amount of
FSWW08 (10, 30, 60 FSWW08) caused a decrease in
the expression of MDR1. 60 FSWW08 had the effect of
a 3-4-fold increase while the available amount of 100
DOX would have caused a 17-fold increase in mono
therapy. Apparently the effect of one of the two agents
on the MDR1 gene prevailed. If Doxorubicin and
FSW008 were given in average concentrations, no
increase in MDR1 gene expression was achieved. This
means that at low physiologic concentrations
fermented soy bean (FSWW08) eliminates drug
resistance caused by doxorubicin.
ERBB2 Gene Expression
Doxorubicin alone caused a continuously
decreasing ERBB2 expression (not shown here, only
reported). While at low concentrations for DOX is still
more than 0.7 times of untreated BT-474, higher DOX
concentrations already down regulates this receptor
tyrosine kinase by 10 times (Table 1).
Estrogen Receptors Alpha and Beta Gene
Estrogen receptor alpha is the classical Estrogen
receptor alpha to describe breast cancer risk. As can
be seen in Table 1, FSWW08 did not change Estrogen
receptor alpha gene expression, but did statistically
significantly increase Estrogen receptor beta gene
expression (Figure 9).
Figure 9: Circulating Tumor Cells (CTCs) of prostate cancer
patients were analyzed for expression of ER-alpha (Estrogen
receptor-alpha) and ER-beta, n = 6 mean and SEM. Asterisk
denotes differences reach statistically significant different on
a 5% probability level.
Gene expression studies with from blood extracted
CTC from cancer patients (Table 1). Table 1 compares
several gene expression investigations obtained by oral
consumption of fermented soy bean FSWW08 after 3
month consumptions with those seen in vitro cell
culture. Estrogen receptor alpha and beta (Figures
9,10): In agreement with in v itro experiments, Estrogen
receptor alpha was not increased in CTC of prostate,
ovarian, and breast cancer, however Estrogen receptor
beta (Table 1).
BAX / Bcl2 Gene Expression
The BAX / Bcl2 ratio is statistically significantly
raised in CTC of prostate cancer patients, however not
in ovarian and breast cancer in vivo (Table 1, Figures
10 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
11-13), suggesting that FSWW08 increases a pro-
apoptotic state of CTC in prostate cancer. BAX was
increased in vitro in breast cancer cells, suggesting a
pro-apoptotic effect as well (Table 1).
Figure 10: Circulating Tumor Cells (CTCs) of ovarian cancer
patients were analyzed for expression of ER-alpha (Estrogen
Receptor-alpha) and ER-beta n=5 (mean and SEM). Asterisk
denotes differences reach statistically significant different on
a 5% probability level.
Figure 11: CTCs (circulating tumor cells) in prostate cancer
patients showed an increased gene expression ratio of
apoptosis-triggering genes Bax/Bcl2 ratio after FSWW08
(fermented soy) therapy.
GAPDH gene expression: All three different CTC
from breast, prostate, and ovarian cancer patients are
decreased (Table 1), although somewhat smaller than
Tp53 Gene Expression
tp53 protein function is considered a gate keeper
and hall mark for cell repair and controls many
important markers in cancer, as well as cell cycle arrest
(Figure 14) and apoptosis. Mutation of tp53 and loss of
gene expression have been detected in many cancers.
It was found that gene expression of tp53 in CTC was
increased in vitro a well as in vivo (Table 1, Figure 15).
The increase in CTC was rather pronounced,
suggesting that in the in vivo case, either additional
mechanisms occur in CTC or fermented soy may
cause additional effects. As is depicted in Figure 15,
tp53 gene expression is sustained over several years.
Tp21 Gene Expression
tp21 is also a key protein in cell repair controlling
apoptosis and cell cycle arrest. It may act together with
Figure 12: CTCs of mama cancer patients showed no
change of gene expression ratio of apoptosis-triggering
genes Bax/Bcl2 after FSWW08 consumption. (Mean and
Standard deviation). Average are not statically significantly
different on a 10% level of significance.
Figure 13: CTCs of breast cancer patients showed no
change of gene expression ratio of apoptosis-triggering
genes bax/bcl2 after 100 days of FSWW08 (fermented soy)
therapy. (Mean and Standard error of the mean; (n = 5).
Mean values are statistically not significant on a 5% level of
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 11
p53 and is important in controlling cancer (Figure 14).
Tp21 is increased rather strongly in vitro as well as in
vivo in CTC. Clearly, the effect of FSWW08 on CTC is
more pronounced than in vitro in all tumors (Table 1,
The expression of the MAPK (Figure 19), was
positively altered by FSWW08 in breast cancer cells.
Additionally reduced inflammation in cancer was
detected with conventional staining technique,
Figure 14: Schematic representation of the relationship between tumor suppressor factors p21 and p53 and their involvement in
apoptosis and cell cycle progression.
Figure 15: Relative gene expression changes of in-blood circulating cancer cells ovarian cancer patients compared to baseline,
compared to expression after 1.5 months, after 3 months and after 4 years (A) prostate cancer patients, (B) breast cancer
patients (C) ovarian cancer patients. Numbers denote individual patients. Taken from ref . All average levels after
consumption reach statistically of significance on the 5 % level of significance.
12 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
corroborating the gene expression results (Figure 20),
where a histological tissue sample of a breast cancer
patient was excised and stained. The conventional
staining technique supports the genetic findings that
immunity is favorably increased in the tumor.
c-Jun Gene Expression
The intracellular release of cytokines in asthma or
allergic reactions is linked to increased c-Jun kinase,
p30 MAP-kinase and NFB in local immune cells .
The first-generation antihistamines have similar
immune modulating responses to FSWW08 and
dexamethasone shares similar ability to alter the gene
expression of p38, MAPkinase, c-Jun and NFB in
myeloma cells, prostate, breast and ovarian cancers
. The reduction of c-Jun in CTC extracted from
Breast tumors supports the findings that immunity is
increased (Figure 19).
It is believed that the decline of TNF alpha in the
blood of patients (Figure 21) is caused by improved
barrier function, so that larger molecules cannot
permeate the cellular barrier (Figure 22). The reduction
in cachexia (Figure 23) by fermented soy bean may
very well be related to improved barrier function,
documented by the improvements of matrix
metalloproteinase 9 gene expression .
Figure 16: Individual relative gene expression changes of tumor suppressor factors p21 in blood CTC (circulating tumor cells),
of prostate cancer patients.
Figure 17: The gene expression of tumor suppressor factors p21 in blood CTC of mamma cancer patients (n=7).
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 13
Figure 18: The individual relative gene expression of tumor suppressor factors p21 in blood CTC extracted form ovarian cancer
Figure 19: Gene expression changes by fermented soy (FSWW08) on blood circulating tumor cancer from breast cancer cells.
The cells were extracted from pooled blood.
VEGF Gene Expression
As is depicted in Figure 24, a statistically significant
increase of VEGF gene expression was detected in
CTC extracted from prostate and ovarian cancer
In cancer patients, appetite and immune status are
significantly weakened. A fermented soy formulation
FSWW08 (group A, consumed FSWW08 orally) and a
placebo formulation (group B) was tested in a clinical
14 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
Figure 20: Histology of a breast cancer sample of a 38-year-old woman. Upper row was taken before consumption and lower
row was taken 14 days later. ‘‘H&E’’ denotes hematoxylin and eosin. Hematoxylin and eosin stain is frequently used for routine
tissue preparation. Eosin is an acid aniline dye. It binds to and stains basic structures (or negatively charged structures), such as
cationic amino groups on proteins. It stains them pink. It can be interpreted as DNA activity and the loss of color as a reduced
DNA activity. F8 as a vascular marker is discussed by Folkman . KI67 is a common marker of anti-angiogenesis. Taken from
Figure 21: TNF-a blood levels before, during, and after daily consumption of fermented soy. Taken from ref .
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 15
Figure 22: A conceptual model of cytokines in cancer. Cancer and immune cells are sources of cytokines, which support the
growth of cancer and lead to psych behavioral symptoms (fatigue, depression, and cognitive impairment), drug toxicity, drug
resistance, anorexia and cachexia, pain, and cancer recurrence and progression. Genetic background, cancer treatment, and
psychological distress can corroborate the production of cytokines. In cancer survivors, hyperactive immune cells might be the
major source of cytokines in psych behavioral symptoms. Taken from ref: .
Figure 23: Influence of daily consumption of a fermented soy formulation on appetite loss (cachexia) in cancer patients under
chemotherapy compared to a group receiving placebo solution containing casein. This was a double-blind study Ref .
study, conducted in six cancer hospitals where cancer
patients underwent radio or chemotherapy (patients
undergoing radiation therapy n = 78, patients
undergoing chemotherapy n = 184, total 262) 
(Figure 23). Group A experienced statistically
significant increases in lymphocyte transformation
rates, whereas group B did not (Figure 25).
Formulations A inhibited or lessened statistically
significant decreases in white blood counts, whereas
the placebo group experienced substantial decreases.
Hemoglobin and platelet decreases were inhibited in
group A, although not statistically significantly .
Patients in group A received no blood transfusions,
whereas many patients from the placebo group
received blood transfusions undergoing chemotherapy
(Figure 24). Appetite loss was reduced in group A from
57.9% to 13.3%. In the placebo group, an increase in
appetite loss was detected under chemo- and radiation
16 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
therapy from 41.8% to 70.9 % (Figure 26). This study
was conducted in China, but similar results were
obtained in the USA at Harvard Medical School .
FSWW08 caused in a small pilot study a higher
survival of ovarian cancer patients compared to
standard therapy, although not statistically significant
. All prostate cancer patients treated with FSWW08
are still alive . The survival of breast cancer
patients increased and the survival rate under
consumption of FSWW08 can be related to breast
cancer patients with high gene expression of tumor
suppressor factor p53, which also show a higher
survival rate than compared to those without it.
The World Health Organization (WHO) reported that
cancer causes 63% of deaths worldwide and is the
second cause of death in western countries .
Cancer is a complex genetic disease with alterations in
multiple cellular signaling pathways . Two traits are
shared in most cancers: genome instability and
mutation tumor-promoting inflammation in the
surrounding tissue so that the tumor can grow into
surrounding tissue . Chemotherapy against the
tumor is mostly conducted with agents inhibiting mitosis
or by other cell toxic mechanisms, for example with
doxorubicin . Only recently it was found that plants
Figure 24: VEGF gene expression in circulating tumor cells extraxted from patients.
Figure 25: Mean and standard deviation of leukocytes before (grey-filled squares) and after (black-filled squares) chemotherapy
with FSWW08, or FSWW08 with yam root extract or a casein placebo solution. Dashed line limits lower range of normal range.
**Denotes a statistically significant difference on the p < 0.01 error level of significance. Taken from ref .
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 17
regulate cell growth similar to animals and humans
. Animals developed later in the evolutionary
development other gene regulating compounds like the
steroidal hormones, where estradiol was the first
steroidal hormone (and was later diversified into the
hormone cascade ).
Plants protect themselves against the growth of a
parasite, what is mechanistically similar to a growth of
a tumor into a surrounding tissue (Figure 26), by DNA
repair mechanism, what is defined as environmental
biotic stress . It is of interest that a tumor may be
mechanistically defined as an unwanted intruder that
imposes stress on its tissue environment to facilitate its
growth (Table 2, Figure 26). Only recently plant
scientists have revealed that many agricultural plants
protect themselves against environmental drought
stress and injuring yeasts or parasite by similar repair
DNA mechanisms, which exist in humans too, like
MAPkinase (Table 2, Figures 4, 19). Hormones and
small RNA actively alter the genomes within a plant
and they have even the capability to conduct this in
other species too, because it was shown that soybean
can pass protective genome mechanism to other plant
species too, raising the question whether this
mechanism can protect against cancer too .
The clinical importance of stress repair mechanism
and the difference to chemotherapy may be illustrated
by the fact that the fermented soy formulation
discussed here in this study caused also a strong and
long lasting reduction of mental stress in soldiers
suffering from PTSD, the posttraumatic stress disorder
: Severe mental stress is a characteristic of PTSD
and cellular and mental stress are related . The
same fermented soy bean formulation has been shown
clinically in humans to stabilize Amyotrophic Lateral
Sclerosis, ALS, what is by definition a progressive cell
disease : Although no complete normalization was
attained, an inhibition of the progressive state was
Figure 26: Schematic comparison of similar growth pattern of a cancer into tissue and a parasite into a leaf.
Table 2: Comparison of cellular gene markers and hormones related to cancer compared to gene described in the
plant as a cell repair gene and determined in plant biology. All genes were determined in soy with the
exception of VEGF like genes and protein. This was found in almost all species with larger tissue besides
yeasts. Foe explanation see text and literature quotation and Figure 4
Biotic Stress Defense
SUPPRESSOR OF GAMMA RESPONSE 1 (SOG 1) Tp53, Tp21 Human Cancer
NO production co ntrol via glutathione peroxidase, phenylalanine-ammonia lyase (PAL), and
PR-1, genes (ref )
IKK/NFB-like signaling pathway
VEGF/PDGF (platelet-derived growth factor) group of the cystine-knot superfamily. (Ref ) VEGF
MAPKinas e  MAPkinas e
Matrix metalloprotein ases ( MMP (ref ) MMP
18 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
seen over several years and for the first time ever. As
was outlined above this plant cell repair mechanism
through abiotic and biotic stress response is non-toxic
and no side effects were reported in cancer multicenter
trial studies and in the ALS study conducted with the
same fermented soy bean formulation, tested here in
vitro gene expression studies.
Clinical improvements in cancer patients, relating to
mental and cellular stress, have been reported
previously by other authors too, investigating the soy
ingredient isoflavone genistein, which resulted in
increased patient survival time, better quality of life,
and reduced adverse effects related to cancer
chemotherapy delivering a dose of 1000 to 1200 mg
Isoflavones a day . We previously reported also
that fermented soy bean (FSWW08), which reduced
stress and improved well-being in soldiers, increased
quality of life in cancer patients as well, reducing
cachexia (Figure 22), reduced side effects of cancer
treatment like cachexia (Figure 23) and leukocytopenia
(Figure 25) and increased the survival time of cancer
A direct comparison of gene expression changes by
FSWW08 formulation was conducted by comparing the
effect on cancer tumor cells (CTC) extracted from
blood of cancer patients to in vitro cancer cell line
treated with cytotoxic doxorubicin (Table 1). Gene
expressions were conducted with the same protocol,
method, and the same laboratory. On the other side
CTC of breast cancer patients and the in vitro breast
cancer cell line are not from the same donor
population. Also, CTC subpopulations might exist with
different properties, because cancer cells having the
ability to form micro metastases and metastases are
likely emerge from CTC, a high medical need exists to
develop new methods to characterize the genotype and
phenotype of CTC . Although we have not
investigated whether subpopulations in our CTC exist,
the alterations by fermented soy bean formulations
Comparisons of gene expression cancer marker by
fermented soy bean in CTC with the breast cancer cell
line BT shows that many anticancer effects like
increase of BAX, increase of tp53, c-Jun, tp21,
silencing of MAPkinase are increased in a similar
manner in vitro cell lines and in CTC (Table 1). This
investigation corroborates other in v itro investigators,
who also saw an improvement of tp53 gene expression
in vitro , as well as MAPkinase , NF-B ,
matrix metalloproteinase [53-55], investigating soy
This here presented investigation is unique since for
the first time ever gene expression in circulating tumor
cells extracted from patient blood are represented and
are additionally related to new findings in the area of
genetic plant science (Table 2), where plants show a
mechanism for cell repair under stress not using any
A very important finding is the simultaneous
increase of tp53 and tp21 gene expression in CTC,
which corroborates the strong apoptotic effect of
fermented soy bean formulation, because tp53 reduces
cell cycling, increases cell repair and diminishes
cellular stress (Figure 14) [56, 58]. Modification and
loss of tp53 has been described as a hall mark for
cancer and missing in about 50% of all cancer patients
. Clearly, there are differences between in vitro and
in vivo: in vivo CTC cells have to be considered to be
less differentiated, so that the increase of tp21 and
tp53 gene expression may have been more
pronounced in vivo (Table 1). Tp53 is related to
increased apoptosis of white blood cell formation too
, which we saw in our clinical experiments (Figure
24). Only recently it was found that plants do express
very similar repair genes to tp53 to overcome
environmental stress .
The additional improvement in barrier function
documented by increase of matrix metalloproteinase
gene by soy by us and others [39, 55] in combination
with strong increase in immunity (Figure 20) may very
well explain the reduction in cancer side effects through
inhibiting cytokine release from the tumor into the blood
(Figure 23) but also modify the cancer stem cells
(CSC) in the tumor, which are believed to drive cancer
growth and metastasis. Improving barrier function of
plants may be an important mechanism by plants to
protect themselves against parasites, injuries, and cuts
and they may not have lost its ability when applied in
A special fermented soy bean FSWW08 did
increase VEGF gene expression in CTC of prostate
and in ovarian cancer patients (Table 1, Figure 24).
Precursor of VEGF have not been identified in yeasts,
but in almost all species, where larger tissue is formed
(Table 2) .
The concept and limitation of “anti-VEGF” therapy in
anticancer treatment is currently strongly debated
among researchers, as it failed in breast cancer and
there may be even an acquired resistance to anti-
VEGF therapy in glioblastoma, which may even
increase the invasiveness of Glioblastoma . VEGF
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 19
is a marker for cellular stress in humans: If oxygen
consumption is low, then VEGF is increased in
pregnancy to promote blood vessel growth and oxygen
supply of the fetus [61-64]. VEGF helps to reduce
cellular stress. As can be seen in Figure 20 where
reduced angiogenesis marker and DNA activity have
been identified in breast cancer tissue after
consumption of the fermented soy bean formulation
FSWW08. The recued angiogenesis and improved
barrier functions preventing the diffusion of cytokines
into the general blood stream (Figure 21), which are
normally responsible for depression, inflammation,
fatigue and most of all for cachexia, the appetite loss
(Figure 22), were also improved (Figure 22).
Both substances, Doxorubicin and fermented soy
bean, had a cytotoxic effect in the human breast cancer
cell line BT-474 (Table 1). Both substances increased
at high doses the expression of MDR1, Doxorubicin
even more. At low physiologic doses, however,
fermented soy bean reduced multiple drug resistance
gene MDR1 even in combination with doxorubicin.
The fact that FSWW08 led to a strong increase in
the expression of the pro-apoptotic gene BAX [64, 65]
(Figure 7) in cancer, in vitro, while the traditional
chemotherapeutic agent Doxorubicin left the
expression of this gene unaltered shows that it might at
least be beneficial to combine FSWW08 with
Doxorubicin for a more effective treatment of cancer. It
is not known why FSWW08 did increase BAX in CTC
extracted from blood of prostate cancer (Figure 11), but
left those extracted from ovarian and breast cancer
unchanged (Figures 12, 13). Programmed cell death to
reduce unnecessary or unwanted cells during
development or disease is similar in plants and animals
[66, 67]. Common pathways in plants and animals
leading to cell death has not been proven [68, 69]. In
plants, the triggering of cell death in response to an
invading pathogen results in the formation of a zone of
dead cells in the vicinity of the infection site . This
killing of host cells is frequently associated with
resistance to further pathogen multiplication and
spread . The tobacco mosaic virus is a model plant
parasite and causes host cells death around a
pathogenic intruder to prevent from further migrating
into the cell. Bcl-2/BAX heterodimers gene controls
death and survival of cells in yeasts and in plants [72,
73] and organizes the protection of a plant against an
intruder [74, 75].
NF-B (nuclear factor kappa B) family transcription
factors are master regulators of immune and
inflammatory processes in response to both injury and
infection . NF-B exists not in plant cells. However
direct precursor cascades were found in plants
regulating NO production and is therefore an important
factor in plant physiology . It was shown by many
investigators that many plants can reduce NF-B in
human tissue and cells, as was shown by fermented
soy beans also .
No clinically relevant health claims can be made
from these in vitro and in vivo studies incorporating
FSWW08, since larger statistically relevant studies
have to be conducted. It may be, however, suggested
that more studies have to be conducted combining
plant based medicine with DNA repair with those
conducting DNA cytotoxic mechanism. The here tested
formulation is not palatable for patients used to
Western food. Also it needs to be investigated, which of
the ingredients besides Isoflavones may also have
caused the gene modifications described in this study.
 Somani SM, JA Romano Jr, editors. Chemical Warfare
Agents: Toxicity at Low Levels. Boca Raton, FL: CRC Press,
 War ing MJ. DNA Modif ication and Cancer. Annual Review of
Biochemistry 1981; 50: 159-192.
 Perez-Tomas R. Multidrug Resistance: Retrospect and
Prospects in Anti-Cancer Drug Treatment. Current Medicinal
Chemistry 2006; 13: 1859-1876.
http://dx.doi.or g/10.2174/09298670 6777585077
 Molnár J, Engi H, Hohmann J, Molnár P, Deli J, Wesolowska
O, Michalak K, Wang Q. Reversal of multidrug resistance by
natural substances from plants. Curr Top Med Chem 2010;
http://dx.doi.or g/10.2174/15680261 0792928103
 Wicha M. Cancer Stem Cells: An Old Idea-A Paradigm Shift.
Cancer Res 2006; 66: 1883.
 Visvader JE, Lindeman GJ. Cancer Stem Cells: Current
Status and Evolving Complexities. Cell Stem Cell 2012; 10:
http://dx .doi.org/10.1016/j.stem.2 012.05 .007
 Sethi S, Li Y, S arkar FH . Regulating miRNA by natural
agents as a new strategy for cancer treat ment. Curr Drug
Targets 2013; 14: 1167-74.
http://dx.doi.or g/10.2174/13894501113149 990189
 Markstein M, Dettorre S, Cho J, Neumüller RA, Craig-Müller
S, Perrimon N. Syste matic screen of chemotherapeutics in
Drosophila stem cell tumors. PNAS 2014; 14: 4530-5.
http://dx.doi.or g/10.1073/pnas.1401 160111
 Singer E, Judkins J, Salomonis N, Matlaf L, Soteropoulos P,
McAllister S, Soroceanu L. Reactive oxygen species-
mediated therapeutic res ponse and resistance in
glioblast oma. Cell Death and D isease 2015; 6.
http://dx.doi.or g/10.1038/cddis.201 4.566
 Macheret M, Halazonet is TD. D NA repl ication stress as a
hallmark of cancer. Ann u Rev Pathol 2015; 10: 425-48.
 (a) Biddlestone J , Bandarra D, Rocha S. The rol e of hypoxia
in inflammatory disease ( Review). Int J Mol Med 2015. doi:
20 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
10.3892/ijmm.2015.2079 (b) Giorgi C, Bonora M, Sorrentin o
G, Missiroli S, Poletti F, Suski JM, Galindo Ramirez F,
Rizzuto R, Di Virgilio F, Zito E, Pandolfi PP, Wieckowski MR,
Mammano F, Del Sal G, Pinton P. p53 at the endoplasmic
reticulum regulates apoptosis in a Ca2+-dependent manner.
Proc Natl Acad Sci USA 2015. pii: 201410723.
 von Stechow L, Typas D, Carreras P uigvert J, Oor t L,
Siddappa R, Pines A, Vrieling H, van de Water B, Mullenders
LH, Danen EH. The E3 ubiquitin ligase ARIH1 pr otects
against genotoxic stress by initiating a 4EHP-mediated
mRNA translation arrest. Mol Cell Biol 2015. pii: MCB.01152-
 Richardson C, Yan S, Vestal CG. Oxidative Stress, Bone
Marrow Failure, and Genome I nstability in Hematopoietic
Stem Cells. Int J Mol Sci 2015; 16(2): 2366-2385. Review.
 Paul S, Amundson SA. Differential Effect of Active Smoking
on Gene Ex pression in Male and Female Smokers. J
Carcinog Mutagen 2014; 5: pii 1 000198.
 Wong EY, Wong SC, Chan CM, Lam EK, Ho LY, Lau CP, A u
TC, Chan AK, Tsang CM, Tsao SW, Lui VW, Chan AT.
TP53-induced glycolysis and apoptosis regulator promotes
proliferation and invasive ness of nasophary ngeal c arcinoma
cells. Oncol Lett 2015; 9(2): 569-574.
 Gu Y, Li P, Peng F, Zhang M, Zhang Y, Liang H, Zhao W, Qi
L, Wang H, Wang C, G uo Z. Autophagy-related prog nostic
signature for br east cancer. Mol Carcinog 2015.
 Rapisarda V, Lor eto C, L edda C, Musumec i G, Bracci M,
Santarelli L, Renis M, Ferrante M, Cardile V. Cytotoxicity,
oxidative stress and genotoxicity induc ed by glass fibers on
human alveolar epithelial cell line A549. Toxicol In Vitro
2015. pii: S0887-2333(14)00250-1.
 Vaubourgeix J, Lin G, Dhar N, Chenouard N, Jiang X, Botella
H, Lupoli T, Mariani O, Yang G, Ouerfelli O, Unser M,
Schnappinger D, McKin ney J, N athan C. Stressed
Mycobacteria Us e the Chaperone ClpB to Sequester
Irreversibly Oxidized Proteins Asymmetrically Within and
Between Cells.Cell Hos t Microb e 2015. pii: S1931-
http://dx.doi.or g/10.1016/j.c hom.2014.12.008
 Ganai AA, Khan AA, Malik ZA, Farooqi H. Genistein
modulates the expr ession of NF-B and MAPK (p-3 8 and
ERK1/2), t hereby a ttenuat ing d-Galactosamine induced
fulminant hepatic failure in Wistar rats. Toxicol Appl
Pharmacol 2015. pii: S0041-008X(15)00022-8.
http://dx.doi.or g/10.1016/j.taap.2015.01. 012
 Fulda S, Kögel D. Cell death by autophagy: emerging
molecular mechanisms and implicat ions for ca ncer ther apy.
 Shaukat Z, Liu D, Hussain R, Khan M, Gregory SL. The Role
of JNK Signalling in Responses to Oxidative DNA Da mage.
Curr Drug Targets 2015. [Epub ahead of print].
 Li Y, Wicha MS, Schwartz SJ, Sun D. Implications of cancer
stem cell t heory for cancer chemoprevention by nat ural
dietary compounds. J Nutr Biochem 2011; 9: 799-806.
 Fehm T, Hoffmann O, Aktas B, Becker S, Solomayer EF,
Wallwiener D, Kimmig R, Kasimir-Bauer S. Detection and
characterization of circulating tumor cells in b lood of primary
breast cancer patients by RT-PCR and comparison to status
of bone marrow disseminated cells. Breast Cancer Research
2009; 11: R59.
 Rohr UD, Gocan AG, Bachg D, Schin dler AE. The effect of
fermented soy (FSWW08) on blood hematology and
cachexia in cancer patients. Horm Mol Biol Clin Invest 2010;
 Atkinson NJ, Urwin PE. The in teraction of plant biotic and
abiotic stresses: from genes to the ﬁeld. Journal of
Experimental B otany 2012; 17: 1-21.
 West CE, Waterworth WM, Sunderland PA, Bray CM.
Arabidopsis DNA double-strand break repair pat hways .
Biochem Soc Trans 2004; 32: 964-966.
http://dx.doi.or g/10.1042/BS T03209 64
 Kozak J, West CE, White C, da Costa-Nunes JA, Angelis KJ.
Rapid repair of DNA double strand breaks in Arabidopsis
thaliana is dependent on proteins involved in chromosome
structure maintenance. DNA Repair (Amst) 2014; 8: 413-419.
http://dx.doi.or g/10.1016/j.dnarep. 2008.1 1.012
 Roy S, Roy CS, Singh SK, Das KP. AtPol a homolog of
mammalian DNA polymerase in Arabidopsis thaliana, is
involved in there pair of UV-B induced DNA damage throu gh
the dark repair pathw ay. Plant Cell Phys iol 2011; 52: 448-
 Roy S. Maintenance of genome stab ility in plants: r epairing
DNA doublw strand breaks and chromatin structure stability.
Frontiers in Plant Science 2014.
 Rodriguez MCS, Petersen M, Mundy J. Mitogen-activ ated
protein kinase signaling in plants. Annual Review of Plant
Biology 2010; 61: 621-649.
 Zhang T, Liu Y, Yang T, Zhan g L, Xu S, Xue L, An L. Diverse
signals converge at MAPK cascades in plant. Plant
Physiolo gy and Biochemistry 2006; 44: 274-283.
 Yoshiyama KO, Sakaguchi K, Kimura S. DNA damage
response in plants: conserved and variable response
compared to animals. Biology (Basel) 2014; 2: 1338-1356.
http://dx.doi.or g/10.3390/bio logy2041338
 Biros E1, Kohút A, Biros I, Kalina I, Bogyiová E, Stubna J. A
link between the p53 germ line poly morphisms and white
blood cells apoptosis in lung cancer patients. Lung Cancer
2002; 35: 231-5.
http://dx.doi.or g/10.1016/S 0169-5002(01)0 0446-9
 Schö nrock N, Exner V, Probst A , Gruissem W, Hennig L.
Functional genomic analysis of CAF-1 mutants in
Arabidopsis thaliana. J Biol Chem 2006; 281: 9560-9568.
http://dx.doi.or g/10.1074/jbc.M513 426200
 Ramirez-Parra E, Gutierrez C. E2F regulates FASCIATA 1, a
chromatin assembly gene whose loss switches on the
endocycle and activates gene expression by changing the
epigenetic status. Plan t Physiol 2007; 144: 105-120.
 Kulcheski FR, de Oliveira LFV, Molina LG, et al. Identiﬁcation
of novel soybean microRNAs involved in abiotic and biotic
stresses. BMC Genomics 2011; 12: 307.
 Rotino GL, Rossoni M , Spinardi A, Bracale M. 2007.
Evaluation of transgenic tomato plants ectopically expressing
the rice Osmyb4 gene. Plant Science 2011; 173: 231-239.
 Hu WS, Lin YM, Ho TJ, Chen RJ, Li YH, Tsai FJ, Tsai CH,
Day CH, Chen TS, Huang CY. Genistein suppresses the
isoproterenol-treated H9c2 cardiomyoblast cell apoptosis
associated with P-38, Erk1/2, JNK, and NFB signaling
protein activation. Am J Chin Med 2013; 41: 1125-36.
 Rohr UD, WW Li, H Ziqiang, Wainright, AE Schindler. The
effect of fer mented soy (FSWW08) on blood hematology a nd
cachexia in cancer patients. Hormone Molecular Biology and
Clinical Investi gation 2012; 12: 407-418.
http://dx.doi.or g/10.1515/hmbci-201 2-0028
 Gocan AG. D Bachg, AE Schindler, UD Rohr. Managing
immunity in resistant cancer patients correlates to survival:
results and discussion of a pilot study. Horm Mol Biol Clin
Invest 20 11; 8: 4 55-469.
 Rostagno P, Moll JL, Bisconte JC, Caldani C. Detection of
Can Plants’ Ability for DNA Repair and Stress Defense Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 21
rare circulating breast cancer cells by filtr ation cytometry and
identific ation by DNA content: sensitivity in an exper iment al
model. Anticancer Res 1997; 17: 2481- 5.
 Hardingham JE, Kotasek D, Farmer B, Butler RN, Mi JX,
Sage RE, Do brovic A. Immuno bead-PCR: a technique for the
detection of circ ulating tumor cells us ing immunoma gnetic
beads and th e polymerase chain reaction. Cancer Res 1993;
 Ferr eira E, Cronjé MJ. Selection of suitable reference genes
for quantitative real-time PCR in apoptosis-induced MCF-7
breast cancer ce lls. Mol Biotec hnol 20 12; 50: 121-12 6.
http://dx.doi.or g/10.1007/s12033-01 1-9425- 3
 Klein A, He X, Roche M, Mallett A, Duska L, Supko JG,
Seiden MV. Prolonged stabilization of platinum-resistant
ovarian cancer in a single patient consuming a fermented soy
therapy. Gynecol Oncol 2006; 100: 205-9.
http://dx.doi.or g/10.1016/j.y gyno.2005.08.006
 Alwan A, Mac Lean DR, Riley LM. Monitoring and
surveillance of chronic non-communicable diseases:
progress and capacity in high- burden countries. The Lancet
2010; 376: 1861-1868.
http://dx.doi.or g/10.1016/S 0140-6736(10)6 1853-3
 Luo J, Solimini NL, Elledge SJ. Pri nciples of cancer therapy:
Oncogene and non-oncogene addiction. Cell 2009; 136(5):
http://dx.doi.or g/10.1016/j.c ell.2009.02.024
 Aggarwal BB, Danda D, Gupta S, Gehlot P. Models for
prevention and treatment of cancer: Pr oblems vs promises.
Biochem Pharmacol 2009; 78: 1083-1094.
 Noolvi MN. Natural sources as potential anti-cancer agents:
A review. International Journal of Phytomedicine 2011; 3: 09-
 Gocan AG, Bachg D, Schindler AE, Rohr UD. Balancing
steroidal hormone cascade in treatment-resistant veteran
soldiers w ith PTSD using a fermented soy prod uct
(FSWW08): a pilot study. Horm Mol Biol Clin Invest 2012; 10:
 Bachen, EA, Manuck SB, Cohen S, Muldoon MF, R aible R,
Herbert TB, Rabin BS. Adrenergic Blockade Ameliorates
Cellular I mmune R esponses to Mental Str ess in Humans.
Psychosomatic Medicine 1995; 57: 366-372.
http://dx.doi.or g/10.1097/00006842-199507 000-00008
 Zhang Z, Wang CZ, Du GJ, Qi Calway LW. Genistein
induces G2/M cell cycle arrest and apoptosis via ATM/p53-
dependen t pathway in human colon cancer cel ls. Int J Oncol
2013; 43: 289-96.
 Bao X, Fen Y, Mian Z, Xue Y, Zhi Z. Effect of genistein on
the TLR and MAPK transduction cascades in
 Sandner A, Illert J, Koitzsch S, Unverzagt S, Schön I. Reflux
induces DNA strand br eaks and expression cha nges of
MMP1+9+1 4 in a human miniorgan culture model. Exp Cell
Res 2013; 319: 2905-15.
http://dx.doi.or g/10.1016/j.y excr.2013.09.004
 Magee PJ, Allsopp P, Samal etdin A, Rowland IR. D aidzein,
R-(+)equol and S-(-)e quol inhibit the invasion of MDA- MB-
231 breast cancer c ells potentially via the dow n-regulation of
matrix metalloproteinase-2. E ur J Nutr 2014; 53: 345-50.
http://dx.doi.or g/10.1007/s00394-01 3-0520-z
 Zheng W, Zh ang Y, Ma D, Shi Y, Liu C, Wang P. (±)Equol
inhibits inv asion in prostate cancer DU145 cells possibly via
down-regulation of matrix metalloproteinase-9, matrix
metalloproteinase-2 and urokinas e-type plasminogen
activaton by antioxidant activity. J Clin Biochem Nutr 201 2;
http://dx.doi.or g/10.3164/jcb n.11-54
 Abbas T, Dutta A. p21 in cancer: intricate networks and
multiple activit ies. Nat Rev C ancer 2009; 9: 400-41 4.
http://dx.doi.or g/10.1038/nrc 2657
 Levine AJ. T he P53 tumor suppressor gene. Na ture 1991;
 Tanner M, Jarvinen P, Isola J. Amplification of HER-2/neu
and topoisomerase IIalph a in primary and metastatic breast
cancer. Cancer R es 2001; 61: 5345-8.
 Piao Y, Liang J, Holmes L, Henry V, Sulman E, de Groot JF.
Acquired resistance to anti-VEGF thera py in glioblastoma is
associated with a mesenchy mal tra nsition. Clin Cancer Res
2013; 19: 4392-4 03.
 Zhao D, Pan C, Sun J, Gilbert C, Drews-Elger K, Azzam DJ,
Picon-Ruiz M, Kim M, Ullmer W, El-Ashry D, Creighton CJ,
Slingerland JM. VEGF drives cancer-i nitiating ste m cells
through VEGFR-2/Stat3 signaling to upregulate Myc and
Sox2. Oncogene 2014.
 Chedraui P, Solis EJ, Bocci G, Gopal S, Russo E, Escobar
GS, Hidalgo L, Pérez- López FR, Genazzani AR, Mannella P,
Simoncini T. Feto-placental nitric oxide, asymmetric
dimethyl arginine and vascular endothelial growth factor
(VEGF) levels and VEGF gene polymorphisms in severe
preeclampsia. J Matern Fet al Neonatal Med 2013; 26: 226-
 Goel HL, Mercurio AM. VEGF targets the tumour cell. Nature
Reviews C ancer 2013; 1 3: 871-8 82.
http://dx.doi.or g/10.1038/nrc 3627
 Keunen O, Johansson M, Oudin A, Sa nzey M, Rahim SA,
Fack F, Thorsen F, Taxt T, Bartos M, Jirik R, Miletic H, Wang
J, Stieber D, Stuhr L, Moen I, Rygh CB, Bjerkvig R, Niclou
SPAnti-VEG F treatment reduces blood supply and increases
tumor cell invasion i n glioblastoma. Proc Natl Acad Sci 2011;
http://dx.doi.or g/10.1073/pnas.101448010 8
 Krajewski S. et al. Prognostic significance of apoptosis
regulators in breast cancer. Endocr Relat Canc er 1999; 6:
 Ikeguchi MS. et al. Quantitative analysis of expression levels
of BAX, Bcl 2 and survivin in cancer cells during cisplatin
treatment. Oncol Rep 20 02; 9: 1 121-6.
http://dx.doi.or g/10.3892/or.9.5.11 21
 Pennell RI, Lamb C. Plant Cell 1997; 9: 1157-1168.
 Greenberg, JT. Plant Mol Biol 1997; 48: 525-545.
 Heath MC, Eur. J P lant Pathol 1998; 104: 117-124.
http://dx.doi.or g/10.1023/A: 1008645520976
 Gilchrist DG. Annu. Rev. Phytopathol 1998; 36: 393-414.
 Lacomme C, Santa Cruz S. B ax-ind uced cell death in
tobacco is similar t o the hypersensitive response. Pr oc Natl
Acad Sci 1999; 96: 7956-7961.
 Goodman RN, N ovacky AJ. The Hypersensitive Reaction in
Plants to Pathogens. A Resistance Phen omenon 1994 APS
Press, St. Paul, MN.
 Kromer G. Nat Med 1997; 3: 614-620.
http://dx.doi.or g/10.1038/nm0697-61 4
 Oltvai ZN, Milliman CL, Korsmeyer SJ. Cell 1993; 74: 609-
http://dx.doi.or g/10.1016/0092-8674( 93)90509-O
 Zha H, Aime-Sempe C, Sato T, Reed JC. J Biol Chem 1997;
 Simonen M, Keller H, Heim J. Eur J Biochem 1997; 249: 85-
http://dx.doi.or g/10.1111/j.1432-1033.1997.t01-1- 00085.x
 Napetschnig J, Wu H. Molecular Basis of NF-B Signaling.
Biophysics 2013; 42: 443-468.
22 Journal of Pharmacy and Nutrition Sciences, 2015, Vol. 5, No. 3 Volko and Rohr
 Golan-Goldhirsh A, Gopas J. Plant derived inhibitors of NF-
B. Phytochemistry Reviews 2014; 13: 107-121.
http://dx.doi.or g/10.1007/s11101-01 3-9293- 5
 Rouleau M, Marsolais F, Richard M, Nicolle L, Voigt B, Adam
G Varin L. Inactivation of Brassinosteroid Biological Activity
by a Salicy late-inducible Steroid Sulfotransferase from
Brassica napus. The J ournal of Biological Chemistry 199 9;
http://dx.doi.or g/10.1074/jbc.274.3 0.20925
 Thornton. Evolution of vertebrate steroid receptors from an
ancestral estrogen receptor by ligand exploit ation a nd serial
genome expansions. PNAS 2001; 98: 5673-79.
 Holmes DHI, Zachar I. The vascular endothelial growth factor
(VEGF) family: angiogenic factors in health and disease.
Genome Biology 2005; 6: 209.
 Negi S, Kharshiing EV, Sharma R. NO way! Is nitric oxide
level in to mato regu lated by a mammalian IKK/NFB-like
signaling pathway? Plant Signal Behav 2011; 6(7): 1049-
 Goslin K, Schindler AE, Rohr UD. Prolonged Sta bilization of
Amyotrophic Lateral Sclerosis (ALS) with a specially
fermented Soy Product (FSWW08): Case Report and
Discussion. Jour nal of N utriti onal Therapeu tics 2013; 2: 1.