Dual effect of metformin on growth inhibition and oestradiol production in breast cancer cells

Rice,S.; Pellat,L.; Ahmetaga,A.; Bano,G.; Mason,H. D.; Whitehead,S. A.

doi:10.3892/ijmm.2015.2108

Dual effect of metformin on growth inhibition and oestradiol production in breast cancer cells

Authors:
- S. Rice
- L. Pellat
- A. Ahmetaga
- G. Bano
- H. D. Mason
- S. A. Whitehead
View Affiliations

Affiliations: Division of Biomedical Sciences, St. George's University of London, London SW17 0RE, UK, Bromley College of F&HE, Bromley Campus, Rookery Lane, Bromley BR2 8HE, UK, MBBS5 Programme, St. George's University of London, London SW17 0RE, UK, Thomas Addison Unit, St. George's Hospital, Cranmer Terrace, London SW17 0RE, UK
Published online on: February 25, 2015 https://doi.org/10.3892/ijmm.2015.2108
Pages: 1088-1094

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Evidence has been accumulating for a role for metformin in reducing breast cancer risk in post-menopausal women. It inhibits growth of breast cancer cells via several mechanisms, primarily the AMPK/mTOR signalling pathway. Another possible protective mechanism may be the ability of metformin to inhibit aromatase activity. In the present study, we investigated the effects of metformin on the basal growth of MCF-7 cells, after oestradiol (E2) stimulation and after the inhibition of mTOR by rapamycin. Secondly, we investigated the effects of metformin on the activity of a number of steroidogenic enzymes and the mRNA expression of aromatase and steroid sulphatase (STS). High doses of metformin significantly inhibited both basal and oestrogen‑stimulated cell division. Low-dose rapamycin (10-10 M) did not inhibit growth, but the addition of metformin induced a significant reduction in growth. High-dose rapamycin (10-8 M) inhibited growth, and this was further attenuated by the addition of metformin. Exposure to low (10-7 M) and high (10-4 M) doses of metformin for 7-10 days significantly reduced the conversion of androstenedione (ANDRO) and testosterone (TESTO) (both requiring aromatase), but not the conversion of oestrone or oestrone sulphate (ES) via 17β-hydroxysteroid dehydrogenase/sulphatase to E2. This attenuation was via a downregulation in the expression of total aromatase mRNA and promoter II, whilst the expression of sulphatase was unaffected by metformin. In conclusion, plasma levels of metformin have a dual therapeutic action, first by directly inhibiting cell proliferation which can be augmented by rapamycin analogues, and secondly, by inhibiting aromatase activity and reducing the local conversion of androgens to E2.

View Figures

View References

1	Ben Sahra I, Le Marchand-Brustel Y, Tanti JF and Bost F: Metformin in cancer therapy: A new perspective for an old anti-diabetic drug. Mol Cancer Ther. 9:1092–1099. 2010. View Article : Google Scholar : PubMed/NCBI
2	Evans JM, Donnelly LA, Emslie-Smith AM, Alessi DR and Morris AD: Metformin and reduced risk of cancer in diabetic patients. BMJ. 330:1304–1305. 2005. View Article : Google Scholar : PubMed/NCBI
3	Gonzalez-Angulo AM and Meric-Bernstam F: Metformin: A therapeutic opportunity in breast cancer. Clin Cancer Res. 16:1695–1700. 2010. View Article : Google Scholar : PubMed/NCBI
4	Gallagher EJ and LeRoith D: Diabetes, cancer, and metformin: Connections of metabolism and cell proliferation. Ann NY Acad Sci. 1243:54–68. 2011. View Article : Google Scholar
5	Soranna D, Scotti L, Zambon A, Bosetti C, Grassi G, Catapano A, La Vecchia C, Mancia G and Corrao G: Cancer risk associated with use of metformin and sulfonylurea in type 2 diabetes: A meta-analysis. Oncologist. 17:813–822. 2012. View Article : Google Scholar : PubMed/NCBI
6	Col NF, Ochs L, Springmann V, Aragaki AK and Chlebowski RT: Metformin and breast cancer risk: A meta-analysis and critical literature review. Breast Cancer Res Treat. 135:639–646. 2012. View Article : Google Scholar : PubMed/NCBI
7	Zhuang Y and Miskimins WK: Cell cycle arrest in metformin treated breast cancer cells involves activation of AMPK, down-regulation of cyclin D1, and requires p27Kip1 or p21Cip. J Mol Signal. 3:18–29. 2008. View Article : Google Scholar
8	Phoenix KN, Vumbaca F and Claffey KP: Therapeutic metformin/AMPK activation promotes the angiogenic phenotype in the ERalpha negative MDA-MB-435 breast cancer model. Breast Cancer Res Treat. 113:101–111. 2009. View Article : Google Scholar :
9	Alimova IN, Liu B, Fan Z, Edgerton SM, Dillon T, Lind SE and Thor AD: Metformin inhibits breast cancer cell growth, colony formation and induces cell cycle arrest in vitro. Cell Cycle. 8:909–915. 2009. View Article : Google Scholar : PubMed/NCBI
10	Feng YH, Velazquez-Torres G, Gully C, Chen J, Lee MH and Yeung SC: The impact of type 2 diabetes and antidiabetic drugs on cancer cell growth. J Cell Mol Med. 15:825–836. 2011. View Article : Google Scholar
11	Jalving M, Gietema JA, Lefrandt JD, de Jong S, Reyners AK, Gans RO and de Vries EG: Metformin: Taking away the candy for cancer. Eur J Cancer. 46:2369–2380. 2010. View Article : Google Scholar : PubMed/NCBI
12	Brown KA, Samarajeewa NU and Simpson ER: Endocrine-related cancers and the role of AMPK. Mol Cell Endocrinol. 366:170–179. 2013. View Article : Google Scholar
13	Li J, Kim SG and Blenis J: Rapamycin: One drug, many effects. Cell Metab. 19:373–379. 2014. View Article : Google Scholar : PubMed/NCBI
14	Vazquez-Martin A, Oliveras-Ferraros C and Menendez JA: The antidiabetic drug metformin suppresses HER2 (erbB-2) oncoprotein overexpression via inhibition of the mTOR effector p70S6K1 in human breast carcinoma cells. Cell Cycle. 8:88–96. 2009. View Article : Google Scholar
15	Chetrite GS, Cortes-Prieto J, Philippe JC, Wright F and Pasqualini JR: Comparison of estrogen concentrations, estrone sulfatase and aromatase activities in normal, and in cancerous, human breast tissues. J Steroid Biochem Mol Biol. 72:23–27. 2000. View Article : Google Scholar : PubMed/NCBI
16	Agarwal VR, Bulun SE, Leitch M, Rohrich R and Simpson ER: Use of alternative promoters to express the aromatase cytochrome P450 (CYP19) gene in breast adipose tissues of cancer-free and breast cancer patients. J Clin Endocrinol Metab. 81:3843–3849. 1996.PubMed/NCBI
17	Barnadas A, Estévez LG, Lluch-Hernández A, Rodriguez-Lescure A, Rodriguez-Sanchez C and Sanchez-Rovira P: An overview of letrozole in postmenopausal women with hormone-responsive breast cancer. Adv Ther. 28:1045–1058. 2011. View Article : Google Scholar : PubMed/NCBI
18	Cook KL, Shajahan AN and Clarke R: Autophagy and endocrine resistance in breast cancer. Expert Rev Anticancer Ther. 11:1283–1294. 2011. View Article : Google Scholar : PubMed/NCBI
19	Brown KA, Hunger NI, Docanto M and Simpson ER: Metformin inhibits aromatase expression in human breast adipose stromal cells via stimulation of AMP-activated protein kinase. Breast Cancer Res Treat. 123:591–596. 2010. View Article : Google Scholar : PubMed/NCBI
20	Samarajeewa NU, Ham S, Yang F, Simpson ER and Brown KA: Promoter-specific effects of metformin on aromatase transcript expression. Steroids. 76:768–771. 2011. View Article : Google Scholar : PubMed/NCBI
21	Anastasius N, Boston S, Lacey M, Storing N and Whitehead SA: Evidence that low-dose, long-term genistein treatment inhibits oestradiol-stimulated growth in MCF-7 cells by down-regulation of the PI3-kinase/Akt signalling pathway. J Steroid Biochem Mol Biol. 116:50–55. 2009. View Article : Google Scholar : PubMed/NCBI
22	Rice S, Amon A and Whitehead SA: Ethanolic extracts of black cohosh (Actaea racemosa) inhibit growth and oestradiol synthesis from oestrone sulphate in breast cancer cells. Maturitas. 56:359–367. 2007. View Article : Google Scholar
23	Gilling-Smith C, Willis DS, Beard RW and Franks S: Hypersecretion of androstenedione by isolated thecal cells from polycystic ovaries. J Clin Endocrinol Metab. 79:1158–1165. 1994.PubMed/NCBI
24	Willis D, Mason H, Gilling-Smith C and Franks S: Modulation by insulin of follicle-stimulating hormone and luteinizing hormone actions in human granulosa cells of normal and polycystic ovaries. J Clin Endocrinol Metab. 81:302–309. 1996.PubMed/NCBI
25	Rice S, Pellatt L, Ramanathan K, Whitehead SA and Mason HD: Metformin inhibits aromatase via an extracellular signal-regulated kinase-mediated pathway. Endocrinology. 150:4794–4801. 2009. View Article : Google Scholar : PubMed/NCBI
26	Rice S, Elia A, Jawad Z, Pellatt L and Mason HD: Metformin inhibits follicle-stimulating hormone (FSH) action in human granulosa cells: Relevance to polycystic ovary syndrome. J Clin Endocrinol Metab. 98:E1491–E1500. 2013. View Article : Google Scholar : PubMed/NCBI
27	Utsumi T, Yoshimura N, Takeuchi S, Maruta M, Maeda K and Harada N: Elevated steroid sulfatase expression in breast cancers. J Steroid Biochem Mol Biol. 73:141–145. 2000. View Article : Google Scholar : PubMed/NCBI
28	Pfaffl MW: A new mathematical model for relative quantification in real-time RT-PCR. Nucleic Acids Res. 29:e452001. View Article : Google Scholar : PubMed/NCBI
29	Robert F, Fendri S, Hary L, Lacroix C, Andréjak M and Lalau JD: Kinetics of plasma and erythrocyte metformin after acute administration in healthy subjects. Diabetes Metab. 29:279–283. 2003. View Article : Google Scholar : PubMed/NCBI
30	Christensen MM, Brasch-Andersen C, Green H, Nielsen F, Damkier P, Beck-Nielsen H and Brosen K: The pharmacogenetics of metformin and its impact on plasma metformin steady-state levels and glycosylated hemoglobin A1c. Pharmacogenet Genomics. 21:837–850. 2011. View Article : Google Scholar : PubMed/NCBI
31	Jung JW, Park SB, Lee SJ, Seo MS, Trosko JE and Kang KS: Metformin represses self-renewal of the human breast carcinoma stem cells via inhibition of estrogen receptor-mediated OCT4 expression. PLoS One. 6:e280682011. View Article : Google Scholar : PubMed/NCBI
32	Berstein LM, Yue W, Wang JP and Santen RJ: Isolated and combined action of tamoxifen and metformin in wild-type, tamoxifen-resistant, and estrogen-deprived MCF-7 cells. Breast Cancer Res Treat. 128:109–117. 2011. View Article : Google Scholar
33	Ali S and Coombes RC: Endocrine-responsive breast cancer and strategies for combating resistance. Nat Rev Cancer. 2:101–112. 2002. View Article : Google Scholar
34	Seto B: Rapamycin and mTOR: A serendipitous discovery and implications for breast cancer. Clin Transl Med. 1:292012. View Article : Google Scholar
35	Beretta L, Gingras AC, Svitkin YV, Hall MN and Sonenberg N: Rapamycin blocks the phosphorylation of 4E-BP1 and inhibits cap-dependent initiation of translation. EMBO J. 15:658–664. 1996.PubMed/NCBI
36	Dowling RJ, Zakikhani M, Fantus IG, Pollak M and Sonenberg N: Metformin inhibits mammalian target of rapamycin-dependent translation initiation in breast cancer cells. Cancer Res. 67:10804–10812. 2007. View Article : Google Scholar : PubMed/NCBI
37	Zakikhani M, Blouin MJ, Piura E and Pollak MN: Metformin and rapamycin have distinct effects on the AKT pathway and proliferation in breast cancer cells. Breast Cancer Res Treat. 123:271–279. 2010. View Article : Google Scholar : PubMed/NCBI
38	Lalau JD, Lemaire-Hurtel AS and Lacroix C: Establishment of a database of metformin plasma concentrations and erythrocyte levels in normal and emergency situations. Clin Drug Investig. 31:435–438. 2011. View Article : Google Scholar : PubMed/NCBI
39	Campagnoli C and Berrino F: Venturelli E, Abbà C, Biglia N, Brucato T, Cogliati P, Danese S, Donadio M, Zito G, Pasanisi P: Metformin dereases circulating androgen and oestrogen levels in nondiabetic women with breast cancer. Breast Cancer. 13:433–438. 2013. View Article : Google Scholar
40	Mansfield R, Galea R, Brincat M, Hole D and Mason H: Metformin has direct effects on human ovarian steroidogenesis. Fertil Steril. 79:956–962. 2003. View Article : Google Scholar : PubMed/NCBI
41	Foster PA, Chander SK, Newman SP, et al: A new therapeutic strategy against hormone-dependent breast cancer: The preclinical development of a dual aromatase and sulfatase inhibitor. Clin Cancer Res. 14:6469–6477. 2008. View Article : Google Scholar : PubMed/NCBI
42	Aka JA, Mazumdar M and Lin SX: Reductive 17beta-hydroxysteroid dehydrogenases in the sulfatase pathway: Critical in the cell proliferation of breast cancer. Mol Cell Endocrinol. 301:183–190. 2009. View Article : Google Scholar
43	Osborne CK and Schiff R: Mechansims of endocrine resistance in breast cancer. Annu Rev Med. 62:233–247. 2011. View Article : Google Scholar
44	Zhu Z, Jiang W, Thompson MD, McGinley JN and Thompson HJ: Metformin as an energy restriction mimetic agent for breast cancer prevention. J Carcinog. 10:172011. View Article : Google Scholar : PubMed/NCBI