Effect of metformin on the proliferation, migration, and MMP-2 and -9 expression of human umbilical vein endothelial cells

  • Authors:
    • Nilufar Esfahanian
    • Yadollah Shakiba
    • Behroz Nikbin
    • Hamid Soraya
    • Nasrin Maleki-Dizaji
    • Mahmood Ghazi-Khansari
    • Alireza Garjani
  • View Affiliations

  • Published online on: January 12, 2012     https://doi.org/10.3892/mmr.2012.753
  • Pages: 1068-1074
Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )
Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )


Abstract

Recent epidemiological studies have demonstrated that metformin lowers the risk of several types of cancer in diabetic patients. Matrix metalloproteinases (MMPs) play a crucial role in the degradation of the vascular basement membrane extracellular matrix proteins, thereby promoting endothelial cell invasion, migration and angiogenesis in the incidence and progression of tumors. The aim of this study was to investigate the effects of metformin on human umbilical vein endothelial cell (HUVEC) proliferation and migration, as well as on MMP-2 and MMP-9 expression. Cell proliferation was determined by cell counting and MTT colorimetric assays. Cell migration was assessed by the wound repair method. Quantitative real-time reverse transcription PCR was performed to quantify the mRNA expression of MMPs. Metformin at concentrations of 0.5-3.0 mM effectively reduced the number of endothelial cells by 5.5-55%, without being cytotoxic to the cells. Similarly, cell proliferation and migration were markedly inhibited by metformin. In addition, treatment with metformin demonstrated a strong (P<0.001) suppressive effect on the mRNA levels of MMP-2 and -9 in the endothelial cells. The inhibitory effects of metformin on endothelial cell number, migration, and MMP expression were reversed partially by compound C, which is an inhibitor of AMP-activated protein kinase (AMPK). The present study reports that metformin considerably inhibited the proliferation, migration, and MMP-2 and -9 expression of HUVECs, and the effect was partially AMPK-dependent. The obtained findings provide a molecular rationale, whereby metformin can exert anticancer effects.

References

1 

D LiSC YeungMM HassanM KonoplevaJL AbbruzzeseAntidiabetic therapies affect risk of pancreatic cancerGastroenterology137482488200910.1053/j.gastro.2009.04.01319375425

2 

JM EvansLA DonnellyAM Emslie-SmithDR AlessiAD MorrisMetformin and reduced risk of cancer in diabetic patientsBMJ33013041305200510.1136/bmj.38415.708634.F715849206

3 

G LibbyLA DonnellyPT DonnanDR AlessiAD MorrisJM EvansNew users of metformin are at low risk of incident cancer: a cohort study among people with type 2 diabetesDiabetes Care3216201625200910.2337/dc08-217519564453

4 

G ZhouR MyersY LiY ChenX ShenJ Fenyk-MelodyM WuJ VentreT DoebberN FujiiRole of AMP-activated protein kinase in mechanism of metformin actionJ Clin Invest10811671174200110.1172/JCI1350511602624

5 

MR OwenE DoranAP HalestrapEvidence that metformin exerts its anti-diabetic effects through inhibition of complex 1 of the mitochondrial respiratory chainBiochem J15348200010839993

6 

N HayN SonenbergUpstream and downstream of mTORGenes Dev1819261945200410.1101/gad.1212704

7 

CS BeeversF LiL LiuS HuangCurcumin inhibits the mammalian target of rapamycin-mediated signaling pathways in cancer cellsInt J Cancer119757764200610.1002/ijc.2193216550606

8 

SA HawleyJ BoudeauJL ReidKJ MustardL UddTP MakelaDR AlessiDG HardieComplexes between the LKB1 tumor suppressor, STRAD alpha/beta and MO25 alpha/beta are upstream kinases in the AMP-activated protein kinase cascadeJ Biol228200310.1186/1475-4924-2-2814511394

9 

JM LizcanoO GoranssonR TothM DeakNA MorriceJ BoudeauSA HawleyL UddTP MakelaDG HardieLKB1 is a master kinase that activates 13 kinases of the AMPK subfamily, including MARK/PAR-1EMBO J23833843200410.1038/sj.emboj.760011014976552

10 

H JiMR RamseyDN HayesLKB1 modulates lung cancer differentiation and metastasisNature448807810200710.1038/nature0603017676035

11 

J FolkmanPA D'AmoreBlood vessel formation: what is its molecular basis?Cell8711531155199610.1016/S0092-8674(00)81810-38980221

12 

R KalluriBasement membranes: structure, assembly and role in tumour angiogenesisNat Rev Cancer3422433200310.1038/nrc109412778132

13 

TA HaasEndothelial cell regulation of matrix metalloproteinasesCan J Physiol Pharmacol8317200510.1139/y04-120

14 

JP SluijterDP de KleijinG PasterkampVascular remodeling and protease inhibition: bench to bedsideCardiovas Res69595603200610.1016/j.cardiores.2005.11.02616387286

15 

BK TanR AdyaJ ChenS FarhatullahD HeutlingD MitchellH LehnertHS RandevaMetformin decreases angiogenesis via NF-kappaB and Erk1/2/Erk5 pathways by increasing the antiangiogenic thrombospondin-1Cardiovasc Res83566574200910.1093/cvr/cvp13119414528

16 

DO XavierLS AmaralMA GomesMetformin inhibits inflammatory angiogenesis in a murine sponge modelBiomed Pharmacother64220225201010.1016/j.biopha.2009.08.00420053525

17 

NJ LinfordDM Dorsa17beta-estradiol and the phytoestrogen genistein attenuate neuronal apoptosis induced by the endoplasmic reticulum calcium-ATPase inhibitor thapsigarginSteroids6710291040200210.1016/S0039-128X(02)00062-4

18 

SL BowkerSR MajumdarP VeugelersJA JohnsonIncreased cancer-related mortality for patients with type 2 diabetes who use sulfonylureas or insulinDiabetes Care29254258200610.2337/diacare.29.02.06.dc05-1558

19 

CJ CurrieCD PooleEA GaleThe influence of glucose-lowering therapies on cancer risk in type 2 diabetesDiabetologia5217661777200910.1007/s00125-009-1440-619572116

20 

JL WrightJL StanfordMetformin use and prostate cancer in Caucasian men: results from a population-based case-control studyCancer Causes Control2016171622200910.1007/s10552-009-9407-y19653109

21 

RM MemmottJR MercadoCR MaierS KawabataSD FoxPA DennisMetformin prevents tobacco carcinogen - induced lung tumorigenesisCancer Prev Res (Phila)310661076201010.1158/1940-6207.CAPR-10-005520810672

22 

YP HwangHG JeongMetformin blocks migration and invasion of tumour cells by inhibition of matrix metalloproteinase-9 activation through a calcium and protein kinase Ca-dependent pathway: phorbol-12-myristate-13-acetate-induced/extracellular signal-regulated kinase/activator protein-1Br J Pharmacol160119512112010

23 

G SchaferBiguanides. A review of history, pharmacodynamics and therapyDiabete Metab914816319836352352

24 

M StumvollN NurjhanG PerrielloG DaileyJE GerichMetabolic effects of metformin in non-insulin-dependent diabetes mellitusN Engl J Med333550554199510.1056/NEJM1995083133309037623903

25 

RS HundalM KrssakS DufourMechanism by which metformin reduces glucose production in type 2 diabetesDiabetes4920632069200010.2337/diabetes.49.12.206311118008

26 

HS HundalT RamlalR ReyesLA LeiterA KlipCellular mechanism of metformin action involves glucose transporter translocation from an intracellular pool to the plasma membrane in L6 muscle cellsEndocrinology131116511731992

27 

D GaluskaLA NolteJR ZierathH Wallberg-HenrikssonEffect of metformin on insulin-stimulated glucose transport in isolated skeletal muscle obtained from patients with NIDDMDiabetologia37826832199410.1007/BF004043407988785

28 

HZ LinSQ YangC ChuckareeF KuhajdaG RonnetAM DiehlMetformin reverses fatty liver disease in obese, leptin-deficient miceNat Med69981003200010.1038/7969710973319

29 

DG HardieSA HawleyJW ScottAMP-activated protein kinase - development of the energy sensor conceptJ Physiol574715200610.1113/jphysiol.2006.10894416644800

30 

KR LaderouteK AminJM CalaoaganM KnappT LeJ OrdunaM ForetzB Viollet5′-AMP-activated protein kinase (AMPK) is induced by low-oxygen and glucose deprivation conditions found in solid-tumor microenvironmentsMol Cell Biol26533653472006

31 

X LiY HanW PangC LiX XieJY ShyyY ZhuAMP-activated protein kinase promotes the differentiation of endothelial progenitor cellsArterioscler Thromb Vasc Biol2817891795200810.1161/ATVBAHA.108.17245218599796

32 

D NagataM MogiK WalshAMP-activated protein kinase (AMPK) signaling in endothelial cells is essential for angiogenesis in response to hypoxic stressJ Biol Chem2783100031006200310.1074/jbc.M30064320012788940

33 

BA KefasY CaiK KerckhofsZ LingG MartensH HeimbergD PipeleersM Van de CasteeleMetformin-induced stimulation of AMP-activated protein kinase in beta-cells impairs their glucose responsiveness and can lead to apoptosisBiochem Pharmacol68409416200410.1016/j.bcp.2004.04.00315242807

34 

M SaitohK NagaiK NakagawaT YamamuraS YamamotoT NishizakiAdenosine induces apoptosis in the human gastric cancer cells via an intrinsic pathway relevant to activation of AMP-activated protein kinaseBiochem Pharmacol6720052011200410.1016/j.bcp.2004.01.020

35 

R RattanS GiriAK SinghI Singh5-Aminoimidazole-4-carboxamide-1-beta-D-ribofuranoside inhibits cancer cell proliferation in vitro and in vivo via AMP-activated protein kinaseJ Biol Chem2803958239593200510.1074/jbc.M50744320016176927

36 

A IsakovicL HarhajiD StevanovicZ MarkovicM Sumarac-DumanovicV StarcevicD MicicV TrajkovicDual antiglioma action of metformin: cell cycle arrest and mitochondria-dependent apoptosisCell Mol Life Sci6412901302200710.1007/s00018-007-7080-417447005

37 

W ZhouWF HanLE LandreeFatty acid synthase inhibition activates AMP-activated protein kinase in SKOV3 human ovarian cancer cellsCancer Res6729642971200710.1158/0008-5472.CAN-06-343917409402

38 

R OkoshiT OzakiH YamamotoK AndoN KoidaS OnoT KodaT KamijoA NakagawaraH KizakiActivation of AMP-activated protein kinase induces p53-dependent apoptotic cell death in response to energetic stressJ Biol Chem28339793987200810.1074/jbc.M70523220018056705

39 

DR AlessiK SakamotoJR BayascasLKB1-dependent signaling pathwaysAnnu Rev Biochem75137163200610.1146/annurev.biochem.75.103004.14270216756488

40 

MM ShawWK GurrRJ McCrimmonDF SchorderetRS Sherwin5'AMP-activated protein kinase alpha deficiency enhances stress-induced apoptosis in BHK and PC12 cellsJ Cell Mol Med112862982007

41 

MB AntonoffJ D'CunhaTeaching an old drug new tricks: metformin as a targeted therapy for lung cancerSemin Thorac Cardiovasc Surg22195196201010.1053/j.semtcvs.2010.10.01521167452

42 

LQ Hong-BrownCR BrownDS HuberCH LangLopinavir impairs protein synthesis and induces eEF2 phosphorylation via the activation of AMP-activated protein kinaseJ Cell Biochem105814823200810.1002/jcb.2188218712774

43 

JE KimHC ChoiLosartan inhibits vascular smooth muscle cell proliferation through activation of AMP-activated protein kinaseKorean J Physiol Pharmacol14299304201010.4196/kjpp.2010.14.5.29921165328

44 

R HanemaaijerP KoolwijkL le ClercqWJ de VreeVW van HinsberghRegulation of matrix metalloproteinase expression in human vein and microvascular endothelial cells. Effects of tumour necrosis factor alpha, interleukin 1 and phorbol esterBiochem J158038091993

45 

Y MorizaneA ThanosK TakeuchiAMP-activated protein kinase suppresses matix metalloproteinase-9 expression in mouse embryonic fibroblastsJ Biol Chem2861603016038201110.1074/jbc.M110.19939821402702

Related Articles

Journal Cover

April 2012
Volume 5 Issue 4

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Esfahanian, N., Shakiba, Y., Nikbin, B., Soraya, H., Maleki-Dizaji, N., Ghazi-Khansari, M., & Garjani, A. (2012). Effect of metformin on the proliferation, migration, and MMP-2 and -9 expression of human umbilical vein endothelial cells. Molecular Medicine Reports, 5, 1068-1074. https://doi.org/10.3892/mmr.2012.753
MLA
Esfahanian, N., Shakiba, Y., Nikbin, B., Soraya, H., Maleki-Dizaji, N., Ghazi-Khansari, M., Garjani, A."Effect of metformin on the proliferation, migration, and MMP-2 and -9 expression of human umbilical vein endothelial cells". Molecular Medicine Reports 5.4 (2012): 1068-1074.
Chicago
Esfahanian, N., Shakiba, Y., Nikbin, B., Soraya, H., Maleki-Dizaji, N., Ghazi-Khansari, M., Garjani, A."Effect of metformin on the proliferation, migration, and MMP-2 and -9 expression of human umbilical vein endothelial cells". Molecular Medicine Reports 5, no. 4 (2012): 1068-1074. https://doi.org/10.3892/mmr.2012.753