All about beans! Apparently, a diet rich in beans is good enough to fight back on tumor progression and genesis.

1. CELL SIGNALLING PATHWAYS
ASSOCIATED WITH THE
REDUCTION IN MAMMARY
CANCER BURDEN BY DIETARY
COMMON BEAN – Matthews D.
Thompson. et al.; Carcinogenesis,
Vol 33, 2012.
Pranamee Sarma

2. INTRODUCTION
- Phaseolus vulgaris (common, dry beans) and mammary cancer
-It is a well known staple food,
widely available and
affordable
-Average consumption can
reach 150-200g/day, BUT,
typical US consumptions are
below recommended levels*
This paper researches on the cancer risk associated with
bean consumption and thus investigates specific Systemic
factors known to contribute to the survival and proliferation of
cancer cells.
*Please see Notes below.
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3. HYPOTHESIS
- So its Man vs Food; Cancer vs Beans
1) Does consumption of beans, really help in reduction of
cancer risk?
2) What systemic factors does it actually affect?
3) The signaling mechanism initiated by bean consumption is
dose-dependent or even traces of it can activate the
cascade?
- Solving these issues would definitely bring to us a new solution
to not treat mammary cancer (which is under the R&D world
already) but will allow us to maintain our dietary structure to be
‘actually’ healthy.
And we always thought, only running and gyming would keep us fit,
healthy and disease free!!!!
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4. OBJECTIVES OF THE STUDY
-So to know the role of Beans in stimulating the health of our
body; we need to study cancer progression focusing on:
1) Systemic Factors(such as glucose-dependent pathways)
2) Cell Autonomous Mechanisms(cellular energy and
networking pathways)
3) Signaling pathways(such as mTOR; this one is the main
crux of beans and cancer risk) **Note

5. METHODS – A Detailed protocol (for all the
curious heads   )
Step I: Female Sprague-Dawley rats were obtained for this
experiment; at 20 days of age(make a point of this) and
maintained at 22⁰C with 50% relative humidity and a 12h
light/dark cycle
Step II: At 21 days of age(here we go), the rats were injected
with 1-methyl-1-nitrosourea(Carcinogen; cancer causing agent)
Step III: Seven days after the injection, all the rats were
randomized based on diet groups (so that we can finally test
beans against others). Bean was incorporated at 60% wt/wt and
for Dose-Response at 7.5%, 15%, 30% and 60% wt/wt. Fed till
46 days post injection
Step IV: Following an overnight fast, rats were ‘euthanized’ over
a 3 h time interval with Carbon monoxide. Blood was obtained,
plasma was isolated by centrifugation at 1000g for 10mins.

6. Step V: Plasma Glucose, insulin, IGF-1, interleukin-6 and C-
reactive protein (CRP) levels were measured using different
assays, such as ELISA for Insulin, IL-6 and CRP.
Step VI: Cell proliferation and apoptosis was determined using
Ki-67 immunohistochemical staining method for the mammary
tissues isolated from the rats. Corresponding hematoxylin- and
eosin-stained serial sections were acquired using Zeiss Axioskop
II at 400x magnification.
Step VII: Western Blotting of the carcinomas (homogenized in
lysis buffer) was performed. 40 ug of protein lysate per sample
was subjected to 8-16% sodium dodecyl sulfate-PAGE gradient
electrophoresis. After electrophoresis, proteins were transferred
to a nitrocellulos membrane. The levels of Cyclin D1, E2F-1, Rb,
p21, p27, Bcl2, X-linked inhibitor of apoptosis proteins, Bax,
AMPK, pACC, ACC, Akt, Raptor were determined using specific
primary antibodies.
Step VIII: Extraction of plasma metabolites using HPLC-Liquid
chromatography and analysis of them using ANOVA was done.

7. RESULTS
Fig 1: Showing the difference in levels of
factors after and before bean-treatment
e.g: We see a decrease in the levels of Cyclin D1 implying
the fact that ratio of hyperphosphrylated Rb to hypo-
decreases in bean-fed rats compared to control ones.***Note

8. Cont……
Figure 2: Dose-dependent effects of bean feeding on
mTOR signaling components (as studied through the
previous data) – Western Blots
As seen from the above data, at a higher dose there seems
to be a considerable increase in the levels of these
components of the mTOR pathway. This is also confirmed
from the next data.

9. Graph A gives a good summarized dose-dependent
curve showing the 60% wt/wt being the best for
reduction of mammary cancer.****

10. mTOR is a conserved serine/threonine
kinase that integrates external cellular
stimuli with intracellular energy and nutrient-
sensing pathways to regulate cellular
metabolism and growth. (presence of Akt
promotes mTOR’s cell growth activity)
In this paper, two upstream regulators of
mTOR pathway were investigated. AMPK (a
nitrogen containing phytochemical
derivative; BTW Common bean is a rich
source of small nitrogen containing
compounds , so here we go!) and Akt (a
downstream effector of the insulin and IGF-1
receptors and both plasma insulin and IGF-1
were reduced dose-dependently with
increasing dietary bean consumption).
mTOR affects lipid metabolism, lipid is
required for cancer cell proliferation. So
AMPK affects mTOR which decreases the
lipid available and thus reduced Cancer risk
.

11. Important word – AMPK (as it affects mTOR
for a better effect)
Twist being: AMPK normally is activated under cellular stress and
nutrient deprivation.. Hmm.. So that needs more study for the
effect of dietary bean on AMPK. I am presuming that AMPK
activation in our case in through LKB1 factor.

12. THUS,
-Common bean consumption appears to reduce mammary
cancer burden through inducing apoptosis (increased
Bax/Bcl2 ratio, p27, p21 increased levels, etc) and
modifying key metabolic signaling networks linked to cell
growth and survival (mTOR being the main one studied)
-We need to know :
a) How AMPK is actually affected?
b) Does the mTOR pathway have a relation with other
factors such as p27 and Bax/Bcl2?
c) What component of Bean and why only this variety?
d) Has study been done also in Asians, who have a
huge intake of dry beans in their regular diet? (Important
as even the environmental effects and bean quality might
differ and give varied results)
Lets research more and make this world a better place to
enjoy and diseases just a mere thought 
THANK YOU!!!!