Anti-angiogenic Effects of Metformin, an AMPK Activator, on Human Umbilical Vein Endothelial Cells and on Granulation Tissue in Rat

Document Type : Original Article

Authors

1 Department of Pharmacology and Toxicology, Faculty of Pharmacy, Tabriz University of Medical Sciences, Tabriz, Iran

2 Department of Immunology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

3 Department of Pharmacology, School of Medicine, Tehran University of Medical Sciences, Tehran, Iran

Abstract

Objective(s)
Metformin is well known for activation of AMP-activated protein kinase (AMPK). AMPK activation inhibits mammalian target of rapamycin (mTOR) as a key signaling process in cell proliferation. Recent epidemiological studies demonstrate that metformin lowers the risk for several types of cancer in diabetic patients. Concerning the critical role of angiogenesis in the incidence and progression of tumors, we investigated the effect of metformin on human umbilical vein endothelial cells migration, as well as on vascular endothelial growth factor (VEGF) expressions in the cells and also on angiogenesis in air pouch model in rats.
Materials and Methods
A "wound" repair method was used to assess the cell migration (n=6). Real-time PCR was performed to quantify the mRNA expression of VEGF (n=5). In air pouch model, carrageenan was injected into the air pouches on the back of rats (n=6) and following an IV injection of carmine red dye granulomatous tissue was processed for the assessment of the dye content. An ordinary ANOVA with Student-Newman-Keuls post hoc test was used to compare groups.
Results
Metformin (orally, 50mg/kg) significantly (P<0.01) decreased angiogenesis in granulomatous tissue by 34% in pouch-bearing rats. Metformin at concentrations of 0.5-3 mM significantly (P<0.001) inhibited VEGF mRNA expression and endothelial cell migration. The inhibitory effects of metformin on the endothelial cell migration were reversed partially by compound C (P<0.01), an inhibitor of AMPK.
Conclusion
The present study reported that metformin inhibited endothelial cell migration and angiogenesis in vitro and in vivo, and the effect was partially AMPK dependent.

Keywords

References

1. Jalving M, Gietema J, Lefrand J, Jong SD, Reyners AKL, Gans ROB, et al. Metformin: Taking away the candy for cancer? Eur J Cancer 2010; 46:2369-2380.
2. Shirwan NA, Zou MH. AMPK in cardiovascular health and disease. Acta Pharmacol Sin 2010; 31:1075-1084.
3. Bowker SL, Majumdar SR, Veugelers P, Johnson JA. Increased cancer-related mortality for patients with type 2 diabetes who use sulfonylureas or insulin. Diabetes Care 2006; 29:254–258.
4. Evans JM, Donnelly LA, Emslie-Smith AM, Alessi DR, Morris AD. Metformin and reduced risk of cancer in diabetic patients. BMJ 2005; 330:1304–1305.
5. Landman GW, Kleefstra N, van Hateren KJ, Groenier KH, Gans ROB, Bilo HJG. Metformin associated with lower cancer mortality in type 2 diabetes: ZODIAC-16. Diabetes Care 2010; 33:322–326.
6. Xavier DO, Amaral LS, Gomes MA, RochaMA, Campos PR, Cota BD, et al. Metformin inhibits inflammatory angiogenesis in a murine sponge model. Biomed Pharmacother 2010; 64:220–225.
7. Drevs J. VEGF and angiogenesis: implications for breast cancer therapy. EJC supplements 2008; 6:7–13.
8. Ben Sahra I, Laurent K, Loubat A, Giorgetti-peraldi S, Glosetti P, Auberger P, et al. The antidiabetic drug metformin exerts an antitumoral effect in vitro and in vivo through a decrease of cyclin D1 level. Oncogene 2008; 27:3576–3586.
9. ClementsA, Gao B, Yeap SHO, Wong MKY, Ali SS, Gurney H. Metformin in prostate cancer: two for the price of one. Ann Oncol 2011; 22:2556-2560.
10. Phoenix KN, Vumbaca F, Claffey KP. Therapeutic metformin/AMPK activation promotes the angiogenic phenotype in the ERalpha negative MDA-MB-435 breast cancer model. Breast Cancer Res Treat 2009; 113:101–111.
11. Zhao X, Zmijewski JW, Lorne E, Liu G, Park YJ, Tsuruta Y, et al. Activation of AMPK attenuates neutrophil proinflammatory activity and decreases the severity of acute lung injury. Am J Physiol Lung Cell Mol Physiol 2008; 295: 497–504.
12. Alba G, El Bekay R, Alvarez-Maqueda M, Chacon P, Vega A, Monteseirin J, et al. Stimulators of AMP-activated protein kinase inhibit the respiratory burst in human neutrophils. FEBS Lett 2004; 573:219–225.
13. Su RY, Chao Y, Chen TY, Huang DY, Lin WW. 5-Aminoimidazole-4-carboxamide riboside sensitizes TRAIL- and TNF_-induced cytotoxicity in colon cancer cells through AMP-activated protein kinase signaling. Mol Cancer Ther 2007; 6:1562–1571.
14. Ghosh AK, Hirasawa N, Niki H, Ohuchi K. Cyclooxygenase-2-mediated angiogenesis in carrageenin-induced granulation tissue in rats. J Pharmacol Exp Ther 2000; 295:802-809.
15. Tan BK, Adya R, Chen J, Farhatullah S, Heutling D, Mitchell D, et al. Metformin decreases angiogenesis via NF-kB and Erk1/2/Erk5 pathways by increasing antiangiogenic thrombospodin-1. Cardiovasc Res 2009; 83:566–574.
16. Tomimoto A, Endo H, Sugiyama M, Fujisawa T, Hosonok K, Takahaski H, et al. Metformin suppresses intestinal polyp growth in Apcmin/+ mice. Cancer Sci 2008; 99:2136–2141.
17. Alimova IN, Liu B, Fan Z, Edgerton SM, Dillon T, Lind SE, et al. Metformin inhibits breast cancer cell growth, colony formation and induces cell cycle arrest in vitro. Cell cycle2009; 8:909–915.
18. Walczak C, Gaignier F, Gilet A, Zou F, Thornton SN, Ropars A. Aldosterone increases VEGF-A production in human neutrophils through PI3K, ERK1/2 and p38 pathways. Biochim Biophys Acta 2011; 1813:2125-2132. 19. Gerber HP, McMurtrey A, Kowalski J, Yan M, Keyt BA, Dixit V, et al. Vascular endothelial growth factor regulates endothelial cell survival through the phosphatidylinositol 3'-kinase/Akt signal transduction pathway. Requirement for Flk-1/KDR activation. J Biol Chem 1998; 13; 273:30336-30343.
20. Dvorak HF.Vascular permeability factor/Vascular endothelial growth factor: a critical cytokine in tumor angiogenesis and a potential target for diagnosis and therapy. J Clin Oncol 2002; 21:4368–4380.
21. Hattori Y, Suzuki K, Hattori S, Kasai K. Metformin inhibits cytokine-induced nuclear factor kappaB activation Via AMP-activated protein kinase activation in vascular endothelial cells. Hypertension 2006; 47:1183-1188.
22. Zhou G, Myers R, Li Y, Chen Y, Shen X, Fenyk-Melody J, et al. Role of AMP-activated protein kinase in mechanism of metformin action. J Clin Invest 2001; 108:1167-1174.
23. Rattan R, Graham RP, Maguire JL, Giri S, Shridhar V. Metformin suppresses ovarian cancer growth and metastasis with enhancement of cisplatin cytotoxicity in vivo. Neoplasia 2011; 13:483-491.
Iranian Journal of Basic Medical Sciences
Volume 15, Issue 6 - Serial Number 6
November 2012
Pages 1202-1209

Files

Share

How to cite

Statistics