Metformin exerts synergistic anti-proliferative effects with Liraglutide in human umbilical vein endothelial cells (HUVECs)

Document Type : Original Article

Authors

1 Department of Toxicology, Faculty of Medical Sciences, Tarbiat Modares University, Tehran, Iran

2 Department of Medical Biotechnology, Faculty of Allied Medicine, Iran University of Medical Sciences, Tehran, Iran

3 Department of Pharmacognosy and Pharmaceutical Biotechnology, School of Pharmacy, Iran University of Medical Sciences, Tehran, Iran

4 Pharmaceutical Sciences Research Center, Tehran Medical Sciences, Islamic Azad University, Tehran Iran

5 Department of Pharmaceutics, School of Pharmacy, Iran University of Medical Sciences, Tehran, Iran

Abstract

Objective(s): Metformin (Met) and Liraglutide (Lira) have been approved to treat type 2 diabetes mellitus and have cardioprotective effects. 
Materials and Methods: Human umbilical vein endothelial cells (HUVECs) were incubated with Met, Lira, or their combination in this study. 
Results: Results showed that the synergistic inhibitory effect of the two drugs on HUVECs proliferation was significant (75%) after 48 hr drug exposure. In addition, either Lira or Met alone had a marked tendency to inhibit the migration of HUVECs (42% and 39%). Almost a complete inhibition (97%) was demonstrated in combinational use after 48 hr treatment. After combining these two drugs, the apoptosis rate raised to 68%, which was a significant approval of synergistic apoptosis induction of Met and Lira. The combinational group indicated a substantial increase in VEGF, PDGF, and MMP-9 at 24 hr compared with the control. 
Conclusion: This study showed that combination therapy with Lira and Met could effectively reduce cell proliferation, induce apoptosis, and inhibit cell migration in the HUVECs. This study provides evidence to support using Met in combination with Lira as a treatment option for patients with type-2 diabetes and cancer.

Keywords


1. Naoyo N, Hirohisa Y, Takashi N, Toshiharu K, Masamichi K. Angiogenesis in cancer. Vasc Health Risk Manag 2006; 2:213-219.
2. Viallard C, Larrivée B. Tumor angiogenesis and vascular normalization: alternative therapeutic targets. Angiogenesis 2017; 20:409-426.
3. Pearson-Stuttard J, Zhou B, Kontis V, Bentham J, Gunter MJ, Ezzati M. Retracted: Worldwide burden of cancer attributable to diabetes and high body-mass index: a comparative risk assessment. Elsevier; 2018.
4. Zaafar DK, Zaitone SA, Moustafa YM. Role of metformin in suppressing 1, 2-dimethylhydrazine-induced colon cancer in diabetic and non-diabetic mice: effect on tumor angiogenesis and cell proliferation. PloS one 2014; 9.
5. Di Y, He J, Ma P, Shen N, Niu C, Liu X, et al. Liraglutide promotes the angiogenic ability of human umbilical vein endothelial cells through the JAK2/STAT3 signaling pathway. Biochem Biophys Res Commun 2020.
6. Hu Y, Liu J, Wang G, Xu Y. The effects of exenatide and metformin on endothelial function in newly diagnosed type 2 diabetes mellitus patients: a case–control study. Diabetes Ther 2018; 9:1295-1305.
7. Eftekhari S, Montazeri H, Tarighi P. Synergistic anti-tumor effects of Liraglutide, a glucagon-like peptide-1 receptor agonist, along with Docetaxel on LNCaP prostate cancer cell line. Eur J Pharmacol 2020:173102.
8. Tawfik MK, Mohamed MI. Exenatide suppresses 1, 2-dimethylhydrazine-induced colon cancer in diabetic mice: Effect on tumor angiogenesis and cell proliferation. Biomed Pharmacother 2016; 82:106-116.
9. Hwang YP, Jeong HG. Metformin blocks migration and invasion of tumour cells by inhibition of matrix metalloproteinase‐9 activation through a calcium and protein kinase Cα‐dependent pathway: Phorbol‐12‐myristate‐13‐acetate‐induced/extracellular signal‐regulated kinase/activator protein‐1. Br J pharmacol 2010; 160:1195-1211.
10. Kannarkatt J, Alkharabsheh O, Tokala H, Dimitrov NV. Metformin and angiogenesis in cancer-revisited. Oncology 2016; 91:179-184.
11. Qian W, Li J, Chen K, Jiang Z, Cheng L, Zhou C, et al. Metformin suppresses tumor angiogenesis and enhances the chemosensitivity of gemcitabine in a genetically engineered mouse model of pancreatic cancer. Life Sci 2018; 208:253-261.
12. Soraya H, Esfahanian N, Shakiba Y, Ghazi-Khansari M, Nikbin B, Hafezzadeh H, et al. Anti-angiogenic effects of Metformin, an AMPK activator, on human umbilical vein endothelial cells and on granulation tissue in rat. Iran J Basic Med Sci 2012; 15:1202.
13. Wu X, Li S, Xue P, Li Y. Liraglutide, a glucagon-like peptide-1 receptor agonist, facilitates osteogenic proliferation and differentiation in MC3T3-E1 cells through phosphoinositide 3-kinase (PI3K)/protein kinase B (AKT), extracellular signal-related kinase (ERK)1/2, and cAMP/protein kinase A (PKA) signaling pathways involving β-catenin. Exp Cell Res 2017; 360:281-291.
14. Helmstädter J, Frenis K, Filippou K, Grill A, Dib M, Kalinovic S, Pawelke F, et al. Endothelial GLP-1 (Glucagon-Like Peptide-1) receptor mediates cardiovascular protection by liraglutide in mice with experimental arterial hypertension. Arterioscler Thromb Vasc Biol 2020; 40(1):145-158. 
15. Drucker DJ. The cardiovascular biology of glucagon-like peptide-1. Cell Metab 2016 ;24(1):15-30.
16. Gaspari T, Liu H, Welungoda I, Hu Y, Widdop RE, Knudsen LB, et al. A GLP-1 receptor agonist liraglutide inhibits endothelial cell dysfunction and vascular adhesion molecule expression in an ApoE-/- mouse model. Diab Vasc Dis Res 2011;8(2):117-24.
17. Ke J, Wei R, Yu F, Zhang J, Hong T. Liraglutide restores angiogenesis in palmitate-impaired human endothelial cells through PI3K/Akt-Foxo1-GTPCH1 pathway. Peptides 2016; 86:95-101.
18. Lu R, Yang J, Wei R, Ke J, Tian Q, Yu F, et al. Synergistic anti-tumor effects of Liraglutide with Metformin on pancreatic cancer cells. PloS one 2018; 13.
19. Tarfiei GA, Shadboorestan A, Montazeri H, Rahmanian N, Tavosi G, Ghahremani MH, et al. GDF15 induced apoptosis and cytotoxicity in A549 cells depends on TGFBR2 expression. Cell Biochem Funct 2019; 37:320-330.
20. Shadboorestan A, Tarfiei GA, Montazeri H, Sepand MR, Zangooei M, Khedri A, et al. Invasion and migration of MDA‐MB‐231 cells are inhibited by block of AhR and NFAT: role of AhR/NFAT1/β4 integrin signaling. J Appl Toxicol 2019; 39:375-384.
21. Bielenberg DR, Zetter BR. The contribution of angiogenesis to the process of metastasis. Cancer J (Sudbury, Mass) 2015; 21:267-273.
22. Teleanu RI, Chircov C, Grumezescu AM, Teleanu DM. Tumor angiogenesis and anti-angiogenic strategies for cancer treatment. J Clin Med 2019; 9:84.
23. Bisht M, Dhasmana DC, Bist SS. Angiogenesis: Future of pharmacological modulation. Indian J Pharmacol 2010; 42:2-8.
24. Goldberg RB. Cytokine and cytokine-like inflammation markers, endothelial dysfunction, and imbalanced coagulation in development of diabetes and its complications. J Clin Endocrinol Metab 2009; 94:3171-3182.
25. Krasner NM, Ido Y, Ruderman NB, Cacicedo JM. Glucagon-like peptide-1 (GLP-1) analog liraglutide inhibits endothelial cell inflammation through a calcium and AMPK dependent mechanism. PLoS One 2014; 9:e97554.
26. Tomimoto A, Endo H, Sugiyama M, Fujisawa T, Hosono K, Takahashi H, et al. Metformin suppresses intestinal polyp growth in ApcMin/+ mice. Cancer Sci 2008; 99:2136-2141.
27. Kamarudin MNA, Sarker MMR, Zhou J-R, Parhar I. Metformin in colorectal cancer: molecular mechanism, preclinical and clinical aspects. J Exp Clin Cancer Res 2019; 38:491.
28. Langlois A, Mura C, Bietiger W, Seyfritz E, Dollinger C, Peronet C, et al. In vitro and in vivo investigation of the angiogenic effects of liraglutide during islet transplantation. PLoS One 2016; 11:e0147068-e0147068.
29. Arakawa M, Mita T, Azuma K, Ebato C, Goto H, Nomiyama T, et al. Inhibition of monocyte adhesion to endothelial cells and attenuation of atherosclerotic lesion by a glucagon-like peptide-1 receptor agonist, exendin-4. Diabetes 2010; 59:1030-1037.
30. Aronis KN, Chamberland JP, Mantzoros CS. GLP-1 promotes angiogenesis in human endothelial cells in a dose-dependent manner, through the Akt, Src and PKC pathways. Metabolism 2013; 62:1279-1286.
31. Ke J, Liu Y, Yang J, Lu R, Tian Q, Hou W, et al. Synergistic effects of Metformin with Liraglutide against endothelial dysfunction through GLP-1 receptor and PKA signalling pathway. Sci Rep 2017; 7:41085.
32. Ben Sahra I, Laurent K, Loubat A, Giorgetti-Peraldi S, Colosetti 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.
33. Dallaglio K, Bruno A, Cantelmo AR, Esposito AI, Ruggiero L, Orecchioni S, et al. Paradoxic effects of Metformin on endothelial cells and angiogenesis. Carcinogenesis 2014; 35:1055-1066.
34. Esfahanian N, Shakiba Y, Nikbin B, Soraya H, Maleki-Dizaji N, Ghazi-Khansari M, et al. Effect of metformin on the proliferation, migration, and MMP-2 and-9 expression of human umbilical vein endothelial cells. Mol Med Rep 2012; 5:1068-1074.
35. Liu C, Liu Y, He J, Mu R, Di Y, Shen N, et al. Liraglutide increases VEGF expression via CNPY2-PERK pathway induced by hypoxia/reoxygenation injury. Front Pharmacol 2019; 10:789.
36. Albini A, Tosetti F, Li VW, Noonan DM, Li WW. Cancer prevention by targeting angiogenesis. Nat Rev Clin Oncol 2012; 9:498-509.
37. Kishimoto K, Liu S, Tsuji T, Olson KA, Hu G-f. Endogenous angiogenin in endothelial cells is a general requirement for cell proliferation and angiogenesis. Oncogene 2005; 24:445-456.
38. Miyoshi H, Kato K, Iwama H, Maeda E, Sakamoto T, Fujita K, et al. Effect of the anti-diabetic drug metformin in hepatocellular carcinoma in vitro and in vivo. Int J Oncol 2014; 45:322-332.
39. Li S, Hu GF. Angiogenin-mediated rRNA transcription in cancer and neurodegeneration. Int J Biochem Mol Biol 2010; 1:26-35.
40. Wang J-C, Li G-Y, Li P-P, Sun X, Li W-M, Li Y, et al. Suppression of hypoxia-induced excessive angiogenesis by metformin via elevating tumor blood perfusion. Oncotarget 2017; 8:73892-73904.
41. Torres G, Morales PE, García-Miguel M, Norambuena-Soto I, Cartes-Saavedra B, Vidal-Peña G, et al. Glucagon-like peptide-1 inhibits vascular smooth muscle cell dedifferentiation through mitochondrial dynamics regulation. Biochem Pharmacol 2016; 104:52-61.
42. Liu Y, Jiang X, Chen X. Liraglutide and metformin alone or combined therapy for type 2 diabetes patients complicated with coronary artery disease. Lipids Health Dis 2017; 16:227.