DOI QR코드

DOI QR Code

Evaluation of Effects of Metformin in Primary Ovarian Cancer Cells

  • Patel, Seema (Department of Biochemistry, AIIMS) ;
  • Singh, Neeta (Department of Biochemistry, AIIMS) ;
  • Kumar, Lalit (Institute of Rotary and Cancer Hospital (IRCH), AIIMS)
  • Published : 2015.11.04

Abstract

Background: Ovarian cancer is the third most common cause of cancer in Indian women. Despite an initial 70-80% response rate, most patients relapse within 1-2 years and develop chemoresistance. Hence, identification or repositioning of drugs to resensitise ovarian cancer cells to existing chemotherapy is needed. Traditionally immortalized cell lines have been used in research, but these may contain genetic aberrations and chromosomal abnormalities serving as poor indicators of normal cell phenotype and progression of early-stage disease. The use of primary cells, maintained for only short periods of time in vitro, may serve as the best representative for studying in vivo conditions of the tissues from which they are derived. In this study we have attempted to evaluate the effect of metformin (an antidiabetic drug) in primary ovarian cancer cells because of its promising effect in other solid tumours. Materials and Methods: Primary cultures of epithelial ovarian cancer cells established from ascitic fluid of untreated ovarian cancer patients were used. The cells were treated with metformin at doses standardized by MTT assay and its ability to induce apoptosis was studied. The cells were analysed for apoptosis and apoptosis related proteins by flow cytometry and western blotting respectively. Results: Metformin induced apoptosis in ovarian cancer cells, provoking cell cycle arrest in the G0/G1 and S phase. It induced apoptosis in ovarian cancer cells by, down-regulating Bcl-2 and up-regulating Bax expression. Conclusions: Metformin was able to induce apoptosis in primary ovarian cancer cells by modulating the expression of Bcl-2 family proteins. These data are relevant to ongoing translational research efforts exploring the chemotherapeutic potential of metformin.

Keywords

Metformin;ovarian cancer;apoptosis;Bcl 2;Bax

References

  1. Ahmed N, Stenvers KL (2013). Getting to know ovarian cancer ascites: opportunities for targeted therapy-based translational research. Frontiers in Oncology, 3, 256.
  2. Aletti GD, Dowdy SC, Podratz KC, et al (2007). Relationship among surgical complexity, short-term morbidity, and overall survival in primary surgery for advanced ovarian cancer. Am J Obstetrics and Gynecol, 197, 676.
  3. Alimova IN, Liu B, Fan Z, et al (2009). Metformin inhibits breast cancer cell growth, colony formation and induces cell cycle arrest in vitro. Cell Cycle (Georgetown, Tex.), 8, 909-15. https://doi.org/10.4161/cc.8.6.7933
  4. Al-Wahab Z, Mert I, Tebbe C, et al (2015). Metformin prevents aggressive ovarian cancer growth driven by high-energy diet: similarity with calorie restriction. Oncotarget, 6, 10908-23. https://doi.org/10.18632/oncotarget.3434
  5. Arikan SK, Kasap B, Yetimalar H, et al (2014). Impact of prognostic factors on survival rates in patients with ovarian carcinoma. Asian Pac J Cancer Prev, 15, 6087-94. https://doi.org/10.7314/APJCP.2014.15.15.6087
  6. Ben Sahra I, Le Marchand-Brustel Y, Tanti J-F, et al (2010). Metformin in cancer therapy: a new perspective for an old antidiabetic drug? Molecular Cancer Therapeutics, 9, 1092-9. https://doi.org/10.1158/1535-7163.MCT-09-1186
  7. Bian J, Li B, Kou X-J, et al (2014). Clinical applicability of multi-tumor marker protein chips for diagnosing ovarian cancer. Asian Pac J Cancer Prev, 15, 8409-11. https://doi.org/10.7314/APJCP.2014.15.19.8409
  8. Chumworathayi B (2013). Personalized cancer treatment for ovarian cancer. Asian Pac J Cancer Prev, 14, 1661-4. https://doi.org/10.7314/APJCP.2013.14.3.1661
  9. Dunfield LD, Shepherd TG, Nachtigal MW (2002). Primary culture and mRNA analysis of human ovarian cells. Biological Procedures Online, 4, 55-61. https://doi.org/10.1251/bpo34
  10. Emami Riedmaier A, Fisel P, Nies AT, et al (2013). Metformin and cancer: from the old medicine cabinet to pharmacological pitfalls and prospects. Trends in Pharmacological Sciences, 34, 126-35. https://doi.org/10.1016/j.tips.2012.11.005
  11. Gadducci A, Tana R, Landoni F, et al (2013). Analysis of failures and clinical outcome of advanced epithelial ovarian cancer in patients with microscopic residual disease at secondlook reassessment following primary cytoreductive surgery and first-line platinum-based chemotherapy. European J Gynaecological Oncol, 34, 213-7.
  12. Gasparri ML, Attar R, Palaia I, et al (2015). Tumor infiltrating lymphocytes in ovarian cancer. Asian Pac J Cancer Prev, 16, 3635-8. https://doi.org/10.7314/APJCP.2015.16.9.3635
  13. Goodwin PJ, Pritchard KI, Ennis M, et al (2008). Insulinlowering effects of metformin in women with early breast cancer. Clinical Breast Cancer, 8, 501-5. https://doi.org/10.3816/CBC.2008.n.060
  14. Gotlieb WH, Saumet J, Beauchamp M-C, et al (2008). In vitro metformin anti-neoplastic activity in epithelial ovarian cancer. Gynecologic Oncology, 110, 246-50. https://doi.org/10.1016/j.ygyno.2008.04.008
  15. Gupta RK, Banerjee A, Pathak S, et al (2013). Induction of mitochondrial-mediated apoptosis by Morinda citrifolia (Noni) in human cervical cancer cells. Asian Pac J Cancer Prev, 14, 237-42. https://doi.org/10.7314/APJCP.2013.14.1.237
  16. Hanahan D, Weinberg RA (2000). The hallmarks of cancer. Cell, 100, 57-70. https://doi.org/10.1016/S0092-8674(00)81683-9
  17. Hwang IC, Park SM, Shin D, et al (2015). Metformin association with lower prostate cancer recurrence in type 2 diabetes: a systematic review and meta-analysis. Asian Pac J Cancer Prev, 16, 595-600. https://doi.org/10.7314/APJCP.2015.16.2.595
  18. Kirk R (2012). Genetics: Personalized medicine and tumour heterogeneity. Nature Reviews. Clinical Oncology, 9, 250.
  19. Kumar S, Meuter A, Thapa P, et al (2013). Metformin intake is associated with better survival in ovarian cancer: a casecontrol study. Cancer, 119, 555-62. https://doi.org/10.1002/cncr.27706
  20. Lengyel E, Litchfield LM, Mitra AK, et al (2015). Metformin inhibits ovarian cancer growth and increases sensitivity to paclitaxel in mouse models. Am J Obstetrics and Gynecol, 212, 479.
  21. Leone A, Di Gennaro E, Bruzzese F, et al (2014). New perspective for an old antidiabetic drug: metformin as anticancer agent. Cancer Treatment and Research, 159, 355-76. https://doi.org/10.1007/978-3-642-38007-5_21
  22. Lima SCS, Hernandez-Vargas H, Herceg Z (2010). Epigenetic signatures in cancer: Implications for the control of cancer in the clinic. Current Opinion in Molecular Therapeutics, 12, 316-24.
  23. Mhawech-Fauceglia P, Wang D, Samrao D, et al (2014). Clinical Implications of Marker Expression of Carcinoma-Associated Fibroblasts (CAFs) in Patients with Epithelial Ovarian Carcinoma After Treatment with Neoadjuvant Chemotherapy. Cancer Microenvironment, 7, 33-9. https://doi.org/10.1007/s12307-013-0140-4
  24. Mitra A, Mishra L, Li S (2013). Technologies for deriving primary tumor cells for use in personalized cancer therapy. Trends in Biotechnology, 31, 347-54. https://doi.org/10.1016/j.tibtech.2013.03.006
  25. Mitsiades CS, Davies FE, Laubach JP, et al (2011). Future directions of next-generation novel therapies, combination approaches, and the development of personalized medicine in myeloma. J Clin Oncol, 29, 1916-23. https://doi.org/10.1200/JCO.2010.34.0760
  26. Nathan DM, Buse JB, Davidson MB, et al (2009). Medical management of hyperglycemia in type 2 diabetes: a consensus algorithm for the initiation and adjustment of therapy. Diabetes Care, 32, 193-203. https://doi.org/10.2337/dc08-9025
  27. Nevadunsky NS, Van Arsdale A, Strickler HD, et al (2014). Metformin use and endometrial cancer survival. Gynecologic Oncology, 132, 236-40. https://doi.org/10.1016/j.ygyno.2013.10.026
  28. Niranjan B, Lawrence MG, Papargiris MM, et al (2013). Primary culture and propagation of human prostate epithelial cells. Methods in Molecular Biology (Clifton, N.J.), 945, 365-82.
  29. Pollak M (2010). Metformin and other biguanides in oncology: advancing the research agenda. Cancer Prevention Research (Philadelphia, Pa.), 3, 1060-5. https://doi.org/10.1158/1940-6207.CAPR-10-0175
  30. Rai B, Bansal A, Patel FD, et al (2014). Radiotherapy for ovarian cancers - redefining the role. Asian Pac J Cancer Prev, 15, 4759-63. https://doi.org/10.7314/APJCP.2014.15.12.4759
  31. Rattan R, Giri S, Hartmann LC, et al (2011). Metformin attenuates ovarian cancer cell growth in an AMP-kinase dispensable manner. Journal of Cellular and Molecular Medicine, 15, 166-78. https://doi.org/10.1111/j.1582-4934.2009.00954.x
  32. Russell S, Duquette M, Liu J, et al (2015). Combined therapy with thrombospondin-1 type I repeats (3TSR) and chemotherapy induces regression and significantly improves survival in a preclinical model of advanced stage epithelial ovarian cancer. FASEB journal: official publication of the Federation of American Societies for Experimental Biology, 29, 576-88. https://doi.org/10.1096/fj.14-261636
  33. Sakarya DK, Yetimalar MH, Ozbasar D (2015). Novel directions in adjuvant chemotherapy for early stage epithelial ovarian cancer. Asian Pac J Cancer Prev, 16, 4157-60. https://doi.org/10.7314/APJCP.2015.16.10.4157
  34. Shank JJ, Yang K, Ghannam J, et al (2012). Metformin targets ovarian cancer stem cells in vitro and in vivo. Gynecologic Oncology, 127, 390-7. https://doi.org/10.1016/j.ygyno.2012.07.115
  35. Siegel R, Naishadham D, Jemal A (2013). Cancer statistics, 2013. CA: A Cancer Journal for Clinicians, 63, 11-30. https://doi.org/10.3322/caac.21166
  36. Takane H, Shikata E, Otsubo K, et al (2008). Polymorphism in human organic cation transporters and metformin action. Pharmacogenomics, 9, 415-22. https://doi.org/10.2217/14622416.9.4.415
  37. Tebbe C, Chhina J, Dar SA, et al (2014). Metformin limits the adipocyte tumor-promoting effect on ovarian cancer. Oncotarget, 5, 4746-64. https://doi.org/10.18632/oncotarget.2012
  38. Topcul M, Cetin I (2015). Effects of metformin on cell kinetic parameters of MCF-7 breast cancer cells in vitro. Asian Pac J Cancer Prev, 16, 2351-4. https://doi.org/10.7314/APJCP.2015.16.6.2351
  39. Trusheim MR, Burgess B, Hu SX, et al (2011). Quantifying factors for the success of stratified medicine. Nature Reviews. Drug Discovery, 10, 817-33. https://doi.org/10.1038/nrd3557
  40. Tseng C-H (2015). Metformin reduces ovarian cancer risk in Taiwanese women with type 2 diabetes mellitus. Diabetes/Metabolism Research and Reviews.
  41. Wang N-N, Zhao L-J, Wu L-N, et al (2013). Mechanistic analysis of taxol-induced multidrug resistance in an ovarian cancer cell line. Asian Pac J Cancer Prev, 14, 4983-8. https://doi.org/10.7314/APJCP.2013.14.9.4983
  42. Wang X, Wang C, Sun Y-T, et al (2015). Taxol produced from endophytic fungi induces apoptosis in human breast, cervical and ovarian cancer cells. Asian Pac J Cancer Prev, 16, 125-31. https://doi.org/10.7314/APJCP.2015.16.1.125
  43. Xie Y, Wang Y-L, Yu L, et al (2011). Metformin promotes progesterone receptor expression via inhibition of mammalian target of rapamycin (mTOR) in endometrial cancer cells. The Journal of Steroid Biochemistry and Molecular Biology, 126, 113-20. https://doi.org/10.1016/j.jsbmb.2010.12.006
  44. Zakikhani M, Blouin M-J, Piura E, et al (2010). Metformin and rapamycin have distinct effects on the AKT pathway and proliferation in breast cancer cells. Breast Cancer Research and Treatment, 123, 271-9. https://doi.org/10.1007/s10549-010-0763-9
  45. Zhang J-W, Sun Q (2015). Metformin may improve the prognosis of patients with pancreatic cancer. Asian Pac J Cancer Prev, 16, 3937-40. https://doi.org/10.7314/APJCP.2015.16.9.3937
  46. Zhang L-Y, Li P-L, Xu A, et al (2015). Involvement of GRP78 in the resistance of ovarian carcinoma cells to paclitaxel. Asian Pac J Cancer Prev, 16, 3517-22. https://doi.org/10.7314/APJCP.2015.16.8.3517
  47. Zhang Y, Li M-X, Wang H, et al (2015). Metformin downregulates endometrial carcinoma cell secretion of IGF-1 and expression of IGF-1R. Asian Pac J Cancer Prev, 16, 221-5. https://doi.org/10.7314/APJCP.2015.16.1.221

Cited by

  1. Effects of Epothilone A in Combination with the Antidiabetic Drugs Metformin and Sitagliptin in HepG2 Human Hepatocellular Cancer Cells: Role of Transcriptional Factors NF-κB and p53 vol.17, pp.3, 2016, https://doi.org/10.7314/APJCP.2016.17.3.993
  2. Metformin reverses prostate cancer resistance to enzalutamide by targeting TGF-β1/STAT3 axis-regulated EMT vol.8, pp.8, 2017, https://doi.org/10.1038/cddis.2017.417