Effect of metformin on proliferation and apoptosis in human pancreatic cancer cell line CFPAC-1 / 中华胰腺病杂志

ABSTRACT

Objective To investigate the effect of metformin on the proliferation and apoptosis in human pancreatic cancer cell line CFPAC-1,and to explore the potential mechanism.Methods Human pancreatic cancer CFPAC-1 cells were cultured in vitro,and were treated with metformin at different concentrations (1,2.5,5,10,20,40,60 mmol/L) for different durations (24 h,48 h and 72 h),and cells without treatment were considered as control group.Cell proliferation was evaluated by CCK-8,cell cycle was determined by flow cytometry,apoptosis was determined by Annexin V/PI double staining method,and Western blot was used to detect the protein expression of p-AMPK,FAS,cyclin D1,Bcl-xl,Bax,caspase-3 and survivin.Results Metformin could inhibit the proliferation of CFPAC-1 cells in a time and dose dependent manner.Forty-eight hours after 40 mmol/L metformin treatment,the proportion of CFPAC-1 cells in phase G0/G1 was significantly increased [(65.93 ± 0.27)％ vs (42.89± 1.02)％],and the proportion of CFPAC-1 cells in phase G2/M,S was significantly decreased [(22.01 ± 2.95) ％ vs (38.28 ± 4.93) ％,(13.58±0.43)％ vs (20.12 ± 3.38)％],and the difference between the two groups was statistically significant (P ＜0.05).The apoptosis rate was increased from (3.01 ± 0.49) ％ to (32.97 ± 3.19) ％,(P ＜ 0.05) ; and the expression of p-AMPK,Bax,and caspase-3 was increased,while the expression of FAS,cyclin D1,Bcl-xl,survivin were decreased.Conclusions Metformin can inhibit proliferation and promote apoptosis of CFPAC-1 cells mainly by activation of AMPK pathway,and down-regulation of FAS,cyclin D1,survivin and Bcl xl/Bax ratio,as well as up-regulation of caspase-3.