Abstract
Background
Ovarian cancer is one of the most common lethal gynecological malignancies world-wide. Despite an initial 70–80 % response rate, most patients relapse within 1–2 years and develop chemo-resistance. Hence, the identification of novel drugs or the repositioning of known drugs to re-sensitize ovarian cancer cells to existing chemotherapy regimens is needed. Here, we evaluated the effect of metformin (an anti-diabetic drug) on ovarian cancer cells, based on its putative effect on other solid tumors.
Methods
Primary cultures of epithelial ovarian cancer cells established from ascitic fluids of untreated ovarian cancer patients and the SKOV-3 ovarian cancer-derived cell line were used. The respective cells were treated with metformin, carboplatin and paclitaxel alone and its various combinations and their effects, including the ability to induce apoptosis, were examined. Concomitantly, the cells were assessed for the expression of several apoptosis-related mRNAs and proteins using quantitative real time PCR, flowcytometry and Western blotting.
Results
We found that metformin induced apoptosis in the ovarian cancer cells tested, and provoked a cell cycle arrest in the G0/G1 and S-phase. Metformin induced apoptosis by down-regulating Bcl-2 and Bcl-xL expression, and up-regulating Bax and Cytochrome c expression. We also found that the apoptosis induction by metformin could be enhanced by a combinatorial use of carboplatin and/or paclitaxel.
Conclusions
Our data indicate that metformin can induce apoptosis in both primary ovarian cancer cells and in SKOV-3 cells. When metformin was combined with carboplatin or paclitaxel, an increased apoptotic activity was observed, implicating a chemo-adjuvant potential.
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References
J.R. Chien, G. Aletti, D.A. Bell, G.L. Keeney, V. Shridhar, L.C. Hartmann, Molecular pathogenesis and therapeutic targets in epithelial ovarian cancer. J. Cell. Biochem. 102, 1117–1129 (2007)
Y. Li, K. Wang, Y.-Z. Jiang, X.-W. Chang, C.-F. Dai, J. Zheng, 2,3,7,8-tetrachlorodibenzo-p-dioxin (TCDD) inhibits human ovarian cancer cell proliferation. Cell. Oncol. 37, 429–437 (2014)
M. Momeni, T. Kalir, S. Farag, L. Chuang, D. Fishman, D.E. Burstein, Expression of H1.5 and PLZF in granulosa cell tumors and normal ovarian tissues: a short report. Cell. Oncol. 37, 229–234 (2014)
J. Di, T. Duiveman-de Boer, P.L.M. Zusterzeel, C.G. Figdor, L.F.A.G. Massuger, R. Torensma, The stem cell markers Oct4A, Nanog and c-Myc are expressed in ascites cells and tumor tissue of ovarian cancer patients. Cell. Oncol. 36, 363–374 (2013)
I.N. Alimova, B. Liu, Z. Fan, S.M. Edgerton, T. Dillon, S.E. Lind, A.D. Thor, Metformin inhibits breast cancer cell growth, colony formation and induces cell cycle arrest in vitro. Cell Cycle 8, 909–915 (2009)
M. Zakikhani, R. Dowling, I.G. Fantus, N. Sonenberg, M. Pollak, Metformin is an AMP kinase-dependent growth inhibitor for breast cancer cells. Cancer Res. 66, 10269–10273 (2006)
G. Zhou, R. Myers, Y. Li, Y. Chen, X. Shen, J. Fenyk-Melody, M. Wu, J. Ventre, T. Doebber, N. Fujii, N. Musi, M.F. Hirshman, L.J. Goodyear, D.E. Moller, Role of AMP-activated protein kinase in mechanism of metformin action. J. Clin. Invest. 108, 1167–1174 (2001)
S.L. Bowker, S.R. Majumdar, P. Veugelers, J.A. Johnson, Increased cancer-related mortality for patients with type 2 diabetes who use sulfonylureas or insulin. Diabetes Care 29, 254–258 (2006)
G.W.D. Landman, N. Kleefstra, K.J.J. van Hateren, K.H. Groenier, R.O.B. Gans, H.J.G. Bilo, Metformin associated with lower cancer mortality in type 2 diabetes: ZODIAC-16. Diabetes Care 33, 322–326 (2010)
G. Libby, L.A. Donnelly, P.T. Donnan, D.R. Alessi, A.D. Morris, J.M.M. Evans, New users of metformin are at low risk of incident cancer A cohort study among people with type 2 diabetes. Diabetes Care 32, 1620–1625 (2009)
S. Jiralerspong, S.L. Palla, S.H. Giordano, F. Meric-Bernstam, C. Liedtke, C.M. Barnett, L. Hsu, M.-C. Hung, G.N. Hortobagyi, A.M. Gonzalez-Angulo, Metformin and pathologic complete responses to neoadjuvant chemotherapy in diabetic patients with breast cancer. J. Clin. Oncol. 27, 3297–3302 (2009)
A. DeCensi, M. Puntoni, P. Goodwin, M. Cazzaniga, A. Gennari, B. Bonanni, S. Gandini, Metformin and cancer risk in diabetic patients: a systematic review and meta-analysis. Cancer Prev. Res. 3, 1451–1461 (2010)
W.H. Gotlieb, J. Saumet, M.-C. Beauchamp, J. Gu, S. Lau, M.N. Pollak, I. Bruchim, In vitro metformin anti-neoplastic activity in epithelial ovarian cancer. Gynecol. Oncol. 110, 246–250 (2008)
R. Rattan, S. Giri, L.C. Hartmann, V. Shridhar, Metformin attenuates ovarian cancer cell growth in an AMP-kinase dispensable manner. J. Cell. Mol. Med. 15, 166–178 (2011)
R. Rattan, R.P. Graham, J.L. Maguire, S. Giri, V. Shridhar, Metformin suppresses ovarian cancer growth and metastasis with enhancement of cisplatin cytotoxicity in vivo. Neoplasia 13, 483–491 (2011)
C.J. Bailey, R.C. Turner, Metformin. N. Engl. J. Med. 334, 574–579 (1996)
K. Mohankumar, S. Pajaniradje, S. Sridharan, V.K. Singh, L. Ronsard, A.C. Banerjea, B.C. Selvanesan, M.S. Coumar, L. Periyasamy, R. Rajagopalan, Apoptosis induction by an analog of curcumin (BDMC-A) in human laryngeal carcinoma cells through intrinsic and extrinsic pathways. Cell. Oncol. 37, 439–454 (2014)
V.V. Dhawan, G.V. Joshi, A.S. Jain, Y.P. Nikam, R.P. Gude, R. Mulherkar, M.S. Nagarsenker, Apoptosis induction and anti-cancer activity of LeciPlex formulations. Cell. Oncol. 37, 339–351 (2014)
T. Nakaoka, A. Ota, T. Ono, S. Karnan, H. Konishi, A. Furuhashi, Y. Ohmura, Y. Yamada, Y. Hosokawa, Y. Kazaoka, Combined arsenic trioxide-cisplatin treatment enhances apoptosis in oral squamous cell carcinoma cells. Cell. Oncol. 37, 119–129 (2014)
O.N. Ikediobi, H. Davies, G. Bignell, S. Edkins, C. Stevens, S. O’Meara, T. Santarius, T. Avis, S. Barthorpe, L. Brackenbury, G. Buck, A. Butler, J. Clements, J. Cole, E. Dicks, S. Forbes, K. Gray, K. Halliday, R. Harrison, K. Hills, J. Hinton, C. Hunter, A. Jenkinson, D. Jones, V. Kosmidou, R. Lugg, A. Menzies, T. Mironenko, A. Parker, J. Perry, K. Raine, D. Richardson, R. Shepherd, A. Small, R. Smith, H. Solomon, P. Stephens, J. Teague, C. Tofts, J. Varian, T. Webb, S. West, S. Widaa, A. Yates, W. Reinhold, J.N. Weinstein, M.R. Stratton, P.A. Futreal, R. Wooster, Mutation analysis of 24 known cancer genes in the NCI-60 cell line set. Mol. Cancer Ther. 5, 2606–2612 (2006)
L.D. Dunfield, T.G. Shepherd, M.W. Nachtigal, Primary culture and mRNA analysis of human ovarian cells. Biol. Proced. Online. 4, 55–61 (2002)
E.L. Tobinick, The value of drug repositioning in the current pharmaceutical market. Drug News Perspect. 22, 119–125 (2009)
C. Campas, Drug repositioning summit: finding new routes to success. Drug News Perspect. 22, 126–128 (2009)
I. Ben Sahra, K. Laurent, A. Loubat, S. Giorgetti-Peraldi, P. Colosetti, P. Auberger, J.F. Tanti, Y. Le Marchand-Brustel, F. Bost, The antidiabetic drug metformin exerts an antitumoral effect in vitro and in vivo through a decrease of cyclin D1 level. Oncogene 27, 3576–3586 (2008)
M. Buzzai, R.G. Jones, R.K. Amaravadi, J.J. Lum, R.J. DeBerardinis, F. Zhao, B. Viollet, C.B. Thompson, Systemic treatment with the antidiabetic drug metformin selectively impairs p53-deficient tumor cell growth. Cancer Res. 67, 6745–6752 (2007)
K. Kisfalvi, G. Eibl, J. Sinnett-Smith, E. Rozengurt, Metformin disrupts crosstalk between G protein-coupled receptor and insulin receptor signaling systems and inhibits pancreatic cancer growth. Cancer Res. 69, 6539–6545 (2009)
A. Isakovic, L. Harhaji, D. Stevanovic, Z. Markovic, M. Sumarac-Dumanovic, V. Starcevic, D. Micic, V. Trajkovic, Dual antiglioma action of metformin: cell cycle arrest and mitochondria-dependent apoptosis. Cell. Mol. Life Sci. 64, 1290–1302 (2007)
Z. Yuan, K. Cao, C. Lin, L. Li, H.-y. Liu, X.-y. Zhao, L. Liu, H.-x. Deng, J. Li, C.-I. Nie, Y.-Q. Wei, The p53 upregulated modulator of apoptosis (PUMA) chemosensitizes intrinsically resistant ovarian cancer cells to cisplatin by lowering the threshold set by Bcl-x(L) and Mcl-1. Mol. Med. 17, 1262–1274 (2011)
Y. Zhuang, W.K. Miskimins, Cell cycle arrest in Metformin treated breast cancer cells involves activation of AMPK, downregulation of cyclin D1, and requires p27Kip1 or p21Cip1. J. Mol. Signal. 3, 18 (2008)
H. Takane, E. Shikata, K. Otsubo, S. Higuchi, I. Ieiri, Polymorphism in human organic cation transporters and metformin action. Pharmacogenomics 9, 415–422 (2008)
L.-W. Wang, Z.-S. Li, D.-W. Zou, Z.-D. Jin, J. Gao, G.-M. Xu, Metformin induces apoptosis of pancreatic cancer cells. World J. Gastroenterol. 14, 7192–7198 (2008)
B. Martin-Castillo, A. Vazquez-Martin, C. Oliveras-Ferraros, J.A. Menendez, Metformin and cancer: doses, mechanisms and the dandelion and hormetic phenomena. Cell Cycle 9, 1057–1064 (2010)
M.R. Owen, E. Doran, A.P. Halestrap, Evidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondrial respiratory chain. Biochem. J. 348, 607–614 (2000)
Acknowledgments
We gratefully acknowledge all patients and IRCH without whose co-operation this study would have been impossible.
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The authors declare that there are no conflicts of interest.
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Patel, S., Kumar, L. & Singh, N. Metformin and epithelial ovarian cancer therapeutics. Cell Oncol. 38, 365–375 (2015). https://doi.org/10.1007/s13402-015-0235-7
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DOI: https://doi.org/10.1007/s13402-015-0235-7