Impact of vitamin C on genistein-induced cell death in prostate cancer. Toluleke Oloruntobi Famuyiwa 09/20/2015
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1 Impact of vitamin C on genistein-induced cell death in prostate cancer 1 st Electronic Conference on Molecular Science Toluleke Oloruntobi Famuyiwa 09/20/2015
2 Introduction
3 Stages of Prostate Cancer
4 Key statistics of prostate cancer Prostate cancer(pca) is the most common cancer in American men. Estimated 220,800 new cases and 27,540 cancer-related deaths in About 1 in every 7 men will be diagnosed with PCa during a lifetime. About 6 cases in 10 are diagnosed in men aged 65 or older. The average age at the time of diagnosis is about 66.
5 Treatment options of prostate cancer Treatment Conventional Non-conventional Chemotherapy Adjuvant therapy /Combination of two therapies
6 Adjuvant therapy / combination of two therapies
7 Objective The Purpose of this study was to: Investigate and determine the potential therapeutic additive effect of genistein and vitamin C on prostate cancer
8 Null Hypothesis Combine treatment of genistein and vitamin C will not cause more cells to die by apoptosis than genistein only.
9 Materials and methods
10 Pictures of cancer cells after treatment
11 Pictures of cancer cells after treatment
12 Pictures of cancer cells after treatment 24hrs (40uM vitamin C + 40uM genistein
13 MTT Assay result Fig A Fig B IC-50 of vitamin C = 40uM IC-50 of genistein = 28uM Higher concentration of vitamin C is needed to kill 50% of LNCaP cells.
14 NBT Assay result
15 Picture of LNCaP cells few minutes before treatment 4 th day few minutes before treatment with drug Cells were seeded into the 96-well MTP at 100ul on the 3 rd day, cells treated when they reached 80% of confluence on 4 th day and MTT assay performed on the 5 th day.
16 Pictures of LNCaP cells 24hrs after treatment Control Vitamin C at 40uM concentration
17 Pictures of LNCaP cells 24hrs after treatment Gen at 30uM + Vitamin C at 40uM
18 Third set of result 1 st day of cell culture 4 th day of cell culture
19 MTT assay Level of significance = 0.05 and p value of t-test =0.11
20 Fluorescence assay of combination treatment showing qualitative result 80uM genistein + 70uM vitamin C 50uM genistein + 70uM vitamin C
21 Quantitative result of florescence assay Genistein 10uM + vitamin C 40uM Genistein 70uM + vitamin C 40uM Control
22 Quantitative result of florescence assay 10uM genistein single treatment 70uM genistein single reatment
23 Percentage of Apoptosis, Necrotic and Living cells in the treatment groups P value = Level of significance = 0.05
24 Ratio of Apoptosis to Necrosis in each treatment Treatment % Apoptosis % Necrosis % Apoptosis/ %Necrosis Control uM of genistein = 3 70uM of genistein = 1 10uM of genistein + 40uM of vitamin C 30uM of genistein + 40uM of vitamin C 70uM of genistein + 40uM of vitamin C = = 10
25 Conclusion P value of chi-square test = and the level of significance is 0.05 Reject the null hypothesis Fluorescence assay shows different number of apoptotic death at different combination concentrations. Decrease in absorbance reflects a decreased level of intracellular ROS The order of effectiveness of treatment is (Gen 10uM + Vit C 40uM) > ( Gen 30uM + Vit C 40uM) >(Gen 70uM + Vit C 40uM) > > 10uM genistein only > 70uM genistein only.
26 References America Cancer Society (2013), Vitamin C. American Cancer Society Du J, Cullen JJ, Buettner GR. Ascorbic acid: chemistry, biology and the treatment of cancer. Biochim Biophys Acta (2012) 1826(2): Hoffman R (2011). Screening for prostate cancer. N Engl J Med, 365, James Kumi-Diaka, Saheed Oluwasina Oseni, Toluleke Famuyiwa, Rolando Branly( 2015) Therapeutic impact of vitamin C on the anticancer activities of genistein isoflavone in radio sensitized LNCaP prostate cancer. J. Cancer Prevention and Current Research. Accepted Research Article. JCPCR-15-RA Parrow NL, Leshin JA, Levine M. Parenteral ascorbate as a cancer therapeutic: a reassessment based on pharmacokinetics. Antioxid Redox Signal (2013) 19(17): Verim L, Yildirim A, Basok EK, et al (2013). Impact of PSA and DRE on histologic findings at prostate biopsy in Turkish men over 75 years of age. Asian Pac J Cancer Prev, 14, YangY, Karakhanova S,Werner J, Bazhin AV(2013) Reactive oxygen species in cancer biology and anticancer therapy. Curr med chem factor-1/chemokine (C-X-C motif) ligand 12 stimulates human hepatoma cell growth,migration, and invasion. Mol Cancer Res 5(1):21 33.
27 Acknowledgement Department of Biological Sciences of Florida Atlantic University
28
Vitamin C Impact on Genistein-Induced Cell Death in Prostate Cancer
OPEN ACCESS Conference Proceedings Paper 1st International Electronic Conference on Molecular Science: Cell Signaling, Survival and Growth Vitamin C Impact on Genistein-Induced Cell Death in Prostate Cancer
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