All-trans retinoic acid and genistein induce cell apoptosis in OVCAR-3 cells by increasing the P14 tumor suppressor gene
Abstract
In this study, we evaluated the effects of all-trans retinoic acid (ATRA) alone or in combination with genistein (GEN) in p14 tumor suppressor gene and subsequent apoptosis of human ovarian carcinoma cells (OVCAR-3). The cells were treated with ATRA or GEN at concentrations of 50 and 25 μM respectively, either alone or in combination for 24 and 48 h. The cell viability was evaluated using 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay. The percentage of cell apoptosis was determined using flow cytometry and p14 gene expression was measured using real time PCR. The MTT results showed that in both ATRA and GEN treated groups, the cell viabilityviability in group treated for 48 h was significantly lower than group treated for 24 h. The flow cytometry results showed that the percentage of apoptotic cells in groups that treated with ATRA and GEN in combination for 24 h and 48 h was significantly more than all other tested groups. The real time results showed that the mRNA level of p14 in cells treated with both drugs for 48 h was significantly higher than all other groups. In conclusion, we confirm that GEN in combination with ATRA is an effective strategy to up regulate the p14 tumor suppressor gene and induce cell apoptosis in OVCAR-3 cell line.
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Maldonado L, Hoque MO. Epigenomics and ovarian carcinoma. Biomark Med. 2010;4(4):543-570.
Romero I, Bast RC Jr. Minireview: human ovarian cancer: biology, current management, and paths to personalizing therapy. Endocrinology. 2012;153(4):1593-1602.
Russo M, Russo GL, Daglia M, Kasi PD, Ravi S, Nabavi SF, et al. Understanding genistein in cancer: The “good” and the “bad” effects: A review. Food Chem. 2016;196:589-600.
Hashiguchi Y, Tsuda H, Yamamoto K, Inoue T, Ishiko O, Ogita S. Combined analysis of p53 and RB pathways in epithelial ovarian cancer. Hum pathol. 2001;32(9):988-996.
Agrawal A, Yang J, Murphy RF, Agrawal DK. Regulation of the p14ARF-Mdm2-p53 pathway: an overview in breast cancer. Exp Mol Pathol. 2006;81(2):115-122.
Yarbrough WG, Bessho M, Zanation A, Bisi JE, Xiong Y. Human tumor suppressor ARF impedes S-phase progression independent of p53. Cancer Res. 2002;62(4):1171-1177.
Hemmati PG, Gillissen B, von Haefen C, Wendt J, Starck L, Guner D, et al. Adenovirus-mediated overexpression of p14ARF induces p53 and Bax-independent apoptosis. Oncogene. 2002;21(20):3149-3161.
Toyota M, Issa JP. Epigenetic changes in solid and hematopoietic tumors. Semin Oncol. 2005;32(5):521-530.
Heo S-H, Kwak J, Jang KL. All-trans retinoic acid induces p53-depenent apoptosis in human hepatocytes by activating p14 expression via promoter hypomethylation. Cancer letters. 2015;362(1):139-48.
Wang Z, Wang L. DNA methylation and esophageal squamous cell carcinoma: special reference to research in china. Life Sci J. 2006;3(2):1-11.
Ozenne P, Eymin B, Brambilla E, Gazzeri S. The ARF tumor suppressor: structure, functions and status in cancer. Int J Cancer. 2010;127(10):2239-2247.
Vaskivuo L, Rysä J, Koivuperä J, Myllynen P, Vaskivuo T, Chvalova K, et al. Azidothymidine and cisplatin increase p14ARF expression in OVCAR-3 ovarian cancer cell line. Toxicol Appl Pharmacol. 2006;216(1):89-97.
Karabulut B, Karaca B, Varol U, Muslu U, Cakar B, Atmaca H, et al. Enhancing cytotoxic and apoptotic effect in OVCAR-3 and MDAH-2774 cells with all-trans retinoic acid and zoledronic acid: a paradigm of synergistic molecular targeting treatment for ovarian cancer. J Exp Clin Cancer Res. 2010;29:102.
Hansen LA, Sigman CC, Andreola F, Ross SA, Kelloff GJ, De Luca LM. Retinoids in chemoprevention and differentiation therapy. Carcinogenesis. 2000;21(7):1271-1279.
Siddikuzzaman, Guruvayoorappan C, Berlin Grace VM. All trans retinoic acid and cancer. Immunopharmacol Immunotoxicol. 2011;33(2):241-249.
Haupt S, Berger M, Goldberg Z, Haupt Y. Apoptosis-the p53 network. J Cell Sci. 2003;116(Pt 20):4077-4085.
Urvalek A, Laursen KB, Gudas LJ. The roles of retinoic acid and retinoic acid receptors in inducing epigenetic changes. Subcell Biochem. 2014;70:129-149.
Zhou RJ, Yang XQ, Wang D, Zhou Q, Xia L, Li MX, et al. Anti-tumor effects of all-trans retinoic acid are enhanced by genistein. Cell Biochem Biophys. 2012;62(1):177-184.
Mukherjee N, Kumar AP, Ghosh R. DNA methylation and flavonoids in genitourinary cancers. Curr Pharmacol Rep. 2015;1(2):112-120.
Choi EJ, Kim T, Lee MS. Pro-apoptotic effect and cytotoxicity of genistein and genistin in human ovarian cancer SK-OV-3 cells. Life Sci. 2007;80(15):1403-1408.
Connolly RM, Nguyen NK, Sukumar S. Molecular pathways: current role and future directions of the retinoic acid pathway in cancer prevention and treatment. Clin Cancer Res. 2013;19(7):1651-1659.
Duvic M, Hymes K, Heald P, Breneman D, Martin AG, Myskowski P, et al. Bexarotene is effective and safe for treatment of refractory advanced-stage cutaneous T-cell lymphoma: multinational phase II-III trial results. J Clin Oncol. 2001;19(9):2456-2471.
Maeno T, Tanaka T, Sando Y, Suga T, Maeno Y, Nakagawa J, et al. Stimulation of vascular endothelial growth factor gene transcription by all trans retinoic acid through Sp1 and Sp3 sites in human bronchioloalveolar carcinoma cells. Am J Respir Cell Mol Biol. 2002;26(2):246-253.
Nagata C, Inaba S, Kawakami N, Kakizoe T, Shimizu H. Inverse association of soy product intake with serum androgen and estrogen concentrations in Japanese men. Nutr Cancer. 2000;36(1):14-18.
Xu ML, Liu J, Zhu C, Gao Y, Zhao S, Liu W, et al. Interactions between soy isoflavones and other bioactive compounds: a review of their potentially beneficial health effects. Phytochem Rev. 2015;14(3):459-467.
Thasni KA, Rojini G, Rakesh SN, Ratheeshkumar T, Babu MS, Srinivas G, et al. Genistein induces apoptosis in ovarian cancer cells via different molecular pathways depending on Breast Cancer Susceptibility gene-1 (BRCA1) status. Eur J Pharmacol. 2008;588(2-3):158-164.
Hebert JR, Hurley TG, Olendzki BC, Teas J, Ma Y, Hampl JS. Nutritional and socioeconomic factors in relation to prostate cancer mortality: a cross-national study. J Natl Cancer Inst. 1998;90(21):1637-1647.
Fotsis T, Pepper M, Adlercreutz H, Fleischmann G, Hase T, Montesano R, et al. Genistein, a dietary-derived inhibitor of in vitro angiogenesis. Proc Natl Acad Sci U S A. 1993;90(7):2690-2694.
Barnes S. Effect of genistein on in vitro and in vivo models of cancer. J Nutr. 1995;125:777S-783S.
Constantinou A, Kiguchi K, Huberman E. Induction of differentiation and DNA strand breakage in human HL-60 and K-562 leukemia cells by genistein. Cancer Res. 1990;50(9):2618-2624.
Peterson G, Barnes S. Genistein and biochanin A inhibit the growth of human prostate cancer cells but not epidermal growth factor receptor tyrosine autophosphorylation. Prostate. 1993;22(4):335-345.
Peterson G, Barnes S. Genistein inhibits both estrogen and growth factor-stimulated proliferation of human breast cancer cells. Cell Growth Differ.. 1996;7(10):1345-1351.
Buckley AR, Buckley DJ, Gout PW, Liang H, Rao YP, Blake MJ. Inhibition by genistein of prolactin-induced Nb2 lymphoma cell mitogenesis. Mol Cell Endocrinol. 1993;98(1):17-25.
Schweigerer L, Christeleit K, Fleischmann G, Adlercreutz H, Wähälä K, Hase T, et al. Identification in human urine of a natural growth inhibitor for cells derived from solid paediatric tumours. Eur J Clin Invest. 1992;22(4):260-264.
Gossner G, Choi M, Tan L, Fogoros S, Griffith KA, Kuenker M, et al. Genistein-induced apoptosis and autophagocytosis in ovarian cancer cells. Gynecol Oncol. 2007;105(1):23-30.
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