Gillian Chugg
Arleigh Grecco
Title: Anti-cancerous effects of vitamin B-complex versus vitamin B6
Abstract:
The objective of this research is to see if vitamin B6 alone or vitamin B-complex slows the
growth of colorectal cancer cells. This research is significant due to conflicting results in the scientific
world, the fact that little research has been published about B vitamin combinations, and that ten
thousands of people die of colorectal cancer a year. The effects of vitamin B complex and vitamin B6 on
a colorectal cancer cell line (HCT-116) are investigated by treating cell media with the proportionate
amounts of vitamins compared to the amount of vitamins that would be ingested by the average adult.
Data was collected by growing a control cell line, a vitamin B complex treated cell line, and a vitamin B6
treated cell line and taking down the cells approximately every other day to get alive cell counts. As a
result of completing this procedure, we can tentatively conclude that B complex has a negative effect on
the growth (makes the cells grow faster) and B6 has a positive effect (makes the cells grow slower). This
research suggests that for a colorectal cancer patient, taking vitamin B-complex will make the cancer
progress faster, whereas taking vitamin B6 alone will slow down cancer cell growth.
Introduction:
There are eight B complex vitamins: thiamine (vitamin B1), riboflavin (vitamin B2), niacin
(vitamin B3), pantothenic acid (vitamin B5), vitamin B6 (pyridoxine, pyridoxamine, or pyridoxal), biotin
(vitamin B7), folic acid (folate or vitamin B9), and vitamin B12 (cobalamin or cyanocobalamin). The B
complex vitamins are water soluble and often found together in natural food sources. The B vitamins are
important for the maintenance and enhancement of the hair, eyes, skin, and liver. The vitamin B
complex is found in vegetables, meat, seeds, eggs, and poultry. Each individual vitamin in the complex
has been researched. Published research delineates certain characteristics that support the idea that the
complex as a whole aids in the prevention and treatment of cancer.
Thiamine is used to assist in the breakdown of sugars, and it prevents nerve and heart problems.
Thiamine has been shown to combat cancer by suppressing transketolase activity in blood, spleen, and
tumor cells. Transketolase is a key enzyme in the pentose phosphate pathway, which is a pathway
extensively used by tumor cells (Thomas et al., 2007).
Riboflavin is essential for vitamin B6 and folate activity by changing vitamin B6 and folate to
their active form. The American Medical Association completed a 400,000 person population study in
which researchers measured the amount of specific vitamins in the individuals’ blood serum. After
looking at the level of riboflavin, there was no correlation found between the amount of riboflavin in the
blood serum and the chance of getting cancer in five years (Johansson et al., 2010).
Niacin works with coenzyme Poly-ADP-ribose polymerases (PARPs). PARPs function in DNA
repair, stress responses, cell signaling, transcription, regulation, apoptosis, chromatin structure, and cell
differentiation (Hidgon & Drake, 2008). The range of niacin’s functions in cell maintenance, particularly
its role in DNA integrity, suggests that niacin might play a role in cancer prevention.
Gillian Chugg
Arleigh Grecco
Pantothenic acid, a part of coenzyme A, is needed for the production of melatonin. An
antitumor effect was shown on mice that received daily amounts of melatonin (Sharman, E.H., Sharman,
K.G., & Bondy, 2011). A possible way to decrease tumor size and number is to increase the amount of
pantothenic acid, which affects melatonin production. Though pantothenic acid and melatonin’s effects
on tumor growth will not be studied directly in this research, the belief that pantothenic acid is
potentially related to cancer is an important topic to note.
Biotin, through biotinylation and holocarboxylase synthetase (HCS), forms histones which are
important proteins for DNA (Hidgon & Drake, 2008). Because cancer prevention and treatment rely on
keeping DNA integrity, biotin may help prevent or treat cancer through its affect on histones.
The sole job of folate coenzymes is “mediating the transfer of one-carbon units” (Hidgon &
Drake, 2008). Folate coenzymes are important in correctly synthesizing DNA and methylation (Hidgon &
Drake, 2008). An above-median level of folate in the blood serum did not show any correlation with a
decrease in lung cancer alone, though (Johansson et al., 2010). Cornelia Ulrich states that high folate
intake aids in cancer prevention, but folate supplementation may be dangerous when cancer has
already developed (“Folate,” 2007, p. 271). Not only does high folate intake aid in cancer prevention,
low folate intake can actually cause cancer because low folate has a chromosome-breaking effect
(“Folate,” 2007, p. 271) (Fenech, 2011). A current problem related to folate is that the current
recommended daily intake may be too low to prevent chromosomal damage (Fenech, 2011).
Vitamin B12 in the form of methylcobalamin is used by the body for methionine sythase, an
enzyme needed for making methionine. Methionine is needed for normal metabolism and growth.
Vitamin B12 is also involved in the metabolism of every cell in the body, especially in DNA synthesis.
However, the American Medical Association studied the level of vitamin B12 in the body, and no
correlation was shown between amounts of vitamin B12 in blood serum and the chance of getting
cancer in five years (Johansson et al., 2010). Michael Fenech has an opposing view, though. Because
vitamin B12 is an essential cofactor for “the maintenance of methylation patterns in DNA,” which affects
genes and chromosomes, and because irregular methylation pattern maintenance increases the risk for
cancer, Fenech has significant reason to believe that low vitamin B12 can cause cancer. Similarly to
folate, the current recommended daily intake for vitamin B12 may be too low to prevent chromosomal
damage (Fenech, 2011).
The specific preventative and anti-cancerous properties of vitamin B6 and the B complex as a
whole will be studied more directly in this research. Vitamin B6 contributes to the body’s formation of
neurotransmitters, leveling homocysteine levels (Sun, Haven, Tsao, & Wu, 2002), and the synthesis of
serine and tetrahydrofolate to glycine, helping to correctly make DNA (van den Donk, Visker, Harryvan,
Kok, & Kampman, 2006). Neurotransmitters help develop melatonin, of which the anticancerous
properties have been previously identified (Hidgon & Drake, 2008). Vitamin B6, in addition to folate and
vitamin B12, contributes to monitoring the amount of homocysteine in the blood (Sun, Haven, Tsao, &
Wu, 2002). Higher levels of homocysteine have been found in cancer patients, but studies indicate that
higher levels of homocysteine are not a cause of cancer and are only an effect of cancerous cell
proliferation (Sun, Haven, Tsao, & Wu, 2002). Because of this, vitamins that regulate levels of
Gillian Chugg
Arleigh Grecco
homocysteine would be more of a treatment for cancer as opposed to a preventive measure. In the
American Medical Association’s population study, above-median levels of vitamin B6 and methionine
showed a 50 % decrease in lung cancer development within five years (Johansson et al., 2010). The
Antitumor effect of vitamin B6 and its mechanisms review states that vitamin B6 may slow down the
development of tumors by slowing down cell development, free radical damage, and the creation of
new blood vessels (Komatsu, Yanaka, Matsubara, & Kato, 2002). A dose dependent relationship has
been identified in cases when vitamin B6 induces chromosomal breakages; the higher the dose, the
greater chance of breakages (Takeuchi, Antunes, & Takahashi, 2007).
Multiple studies have studied the effects of multiple B vitamins in combination. In the American
Medical Association’s population study, above-median levels of vitamin B6, methionine, and folate
showed a 66 % decrease in developing lung cancer within five years versus only a 50% decrease with
above-median levels of vitamin B6 and methionine (Johansson et al., 2010). A different study was
published that combined CoenzymeQ10, riboflavin and niacin (Premkumar, Yuvaraj, Sathish, Shanthi, &
Sachdanandam, 2008). After giving breast cancer patients, CoenzymeQ10, riboflavin and niacin, there
was a decrease in angiogenic influences and an increase in anti-angiogenic influences (Premkumar, et
al., 2008). Studies demonstrate that combinations of B vitamins produce more positive results.
Research has been published on individual components of B vitamins and certain combinations
of different B vitamins together and how preventive or anti-cancerous the vitamins are, however, these
researches could not find studies done with the B complex as a whole in relation to cancer. In addition,
there are multiple contradicting studies. Van den Donk et al. (2006) cites multiple studies that contradict
each other surrounding the B vitamins and cancer, thus concluding that more research is necessary. This
research will look into lung cancer and study how anti-cancerous Vitamin B complex is compared to
Vitamin B6 alone. Previous research suggests that our study may be fulfilling a vitamin B6 deficiency for
cancer patients (Depeint, Bruce, Shangari, Mehta, & O’Brien, 2006). The B vitamins play an essential role
in maintaining mitochondrial function and mitochondria are compromised by a deficiency of any B
vitamin (Depeint et al., 2006). Vitamin B therapy has been shown to alleviate B deficiency symptoms and
prevents mitochondrial toxicity (Depeint et al., 2006). Depeint et al. (2006) found B vitamins to be
effective at preventing oxidative stress toxicity as well as Takeuchi, Antunes, & Takahashi (2007, pg.
669), who found that a vitamin B6 deficiency “decreases the antioxidant defense system” and “increases
oxidative stress in rat liver tissue.” A weak antioxidant defense system and increased oxidative stress are
both qualities common among cancer patients. According to the American Medical Association’s study,
“lung cancer remains the most common cause of cancer death… today” (Johansson et al., 2010, pg.
2384). If more research is done on the anti-cancerous properties of B vitamins, then it helps save or
prolong the lives of those living with lung cancer.
Gillian Chugg
Arleigh Grecco
Methods and Materials:
Basic materials needed for cell culture were acquired and HCT-116 colorectal cancer cells were
cultured in a 75cc flask. Once this flask reached a confluency of approximately 70%, the cells were split
into a well plate with approximately 4,000 cells per well. All of the wells either contained untreated
control media, vitamin B6 (GNC, 255214) treated media, or vitamin B-complex (GNC, 017913)treated
media. The amount of vitamins to be added to the experimental media was determined by a proportion
of the amount of vitamins that would be absorbed by a human body of approximately 5,600mL of blood
proportionate to 4mL of media; the amount of media in each well. Rows of three wells were taken down
on a set schedule of approximately every other day. A hemocytometer was then used to count the alive
cells in each well.
Data:
Table 1: Control trial data of number of alive cells/mL counted on certain days since seeding
Days Since Number of Alive
Seeding Cells/mL
0
4000
4
45185
5
309894
6
757500
7
1795000
8
2276250
Table 2: Vitamin B6 trial data of number of alive cells/mL counted on certain days since seeding
Days Since Number of
Seeding
Alive Cells/mL
0
4000
4
84444.44
6
983750
8
1790000
Gillian Chugg
Arleigh Grecco
Table 3: Vitamin B-complex trial data of number of alive cells/mL counted on certain days since seeding
Days Since Number of
Seeding
Alive Cells/mL
0
4000
4
135000
5
402118
6
1133333
7
1842222
8
2363667
Results:
Table 4: χ2 Values for Vitamin treatment data compared to control data per day
Vitamin B6 χ2 Value Vitamin B-Complex χ2 Value
Day 6
240082.56
831671.24
Day 7
N/A
29811.1
Day 8
721684.43
85145.07
Gillian Chugg
Arleigh Grecco
Graph 1: Compiled Data for Control and
Experimental Trials Comparing Days Since
Seeding and the Number of Alive Cells/mL
2500000
Number of Alive Cells/mL
2000000
1500000
Control
1000000
Vitamin B6 Experimental
Vitamin B-Complex
Experimental
500000
0
0
-500000
2
4
6
Days Since Seeding
8
10
Gillian Chugg
Arleigh Grecco
Discussion:
As a result of completing the stated procedure with multiple trials, the data suggests that Bcomplex has a negative effect on the growth (makes the cells grow faster) and B6 has a positive effect
(makes the cells grow slower). Consequently, the original hypothesis, “If vitamin B6 and vitamin Bcomplex are applied to separate cell cultures of colorectal cancer cells, then the cells applied with the
vitamin B-complex will better slow the growth of the cancer cells because all parts of the B-complex
have anti-cancerous properties that often work synergistically,” is not supported.
With respect to previous research on this topic, these results further support that vitamin B6
has beneficial anti-cancerous properties on its own. Because research on the entire B-complex in
accordance with cancer was unprecedented before this research, these results suggest that all of the B
vitamins used together in accordance with cancer has adverse effects. These conclusions are able to be
drawn because results of the chi-square test that was run on the day six, seven, and eight data points (in
Table 4). The critical value for a 95% confidence interval with two degrees of freedom is 5.99. As seen in
Table 4, all of the calculated chi-square values (240082.56, 721684.43, 831671.24, 29811.1, and
85145.07) far exceed 5.99 indicating statistically significant differences for both the B-complex and B6
treated trials, however the B-complex treated trial is significantly higher and the B6 treated trial is
significantly lower when compared to the control. Nonetheless, this research is still extremely
important. The results of this research cannot be fully compared to how the vitamins react in a human
body system, but the research suggests that a colorectal cancer patient should not take vitamin Bcomplex because B-complex could make the cancer progress faster, but a colorectal cancer patient
should take vitamin B6 alone because vitamin B6 could slow down cancer cell growth.
Future work that can be done in accordance with this project includes testing B-complex and B6
on a different cancer cell line, determining why exactly B6 decreases the growth of colorectal cancer
cells, optimizing vitamin concentrations, or testing different vitamin combinations.
Gillian Chugg
Arleigh Grecco
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Arleigh Grecco
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