二甲双胍对人胰腺癌细胞株CFPAC-1增殖与凋亡的影响

目的观察二甲双胍对人胰腺癌细胞株CFPAC-1增殖、凋亡的影响，探讨其相关分子机制。方法应用1、2．5、5、10、20、40、60mmol／L二甲双胍处理人胰腺癌CFPAC-1细胞24、48、72h，以未处理的细胞作为对照组。采用CCK-8法检测细胞的增殖抑制率，流式细胞仪分析细胞周期，AnnexinV／PI双染法检测细胞凋亡，蛋白质印迹法检测磷酸化AMPK（p-AMPK）、FAS、cyclinD1、Bcl-xl、Bax、caspase-3、survivin蛋白的表达。结果二甲双胍呈浓度及时间依赖性抑制CFPAC-1细胞的增殖。40mmol／L二甲双胍处理细胞48h后，G0／G1细胞比例显著增多[（65．93±0.27）％比（42．89±1．02）％]，G2／M期、s期细胞比例显著减少[（22．01±2．95）％比（38．28±4．93）％，（13．58±0．43）％比（20．12±3．38）％]，差异均有统计学意义（P值均〈0．05）；细胞凋亡率从对照组的（3．01±0．49）％增加到（32．97±3．19）％（P〈0．05）；p-AMPK、Bax、caspase-3表达增强，FAS、cyclinD1、Bcl-xl、survivin表达减弱。结论二甲双胍可以显著抑制CFPAC-1细胞增殖，促进细胞凋亡，其机制可能通过激活AMPK信号通路，下调FAS、cyclinD1、survivin表达及Bcl-xl／Bax比值，上调caspase-3表达所致。

Effect of metformin on proliferation and apoptosis in human pancreatic cancer cell line CFPAC-1

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 Mefformin 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 U （ 65.93 ±0. 27 ） % vs （42.89± 1.02 ） % ] , and the proportion of CFPAC-1 cells in phase GJM, 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 , Bel-xl, survivin were decreased. Conclusions Mefformin can inhibit proliferation and promote apoptosis of CFPAC-1 cells mainly by activation of AMPK pathway, and down-regnlation of FAS, cyclin D1 ,survivin and Bcl xl/Bax ratio, as well as up-regulation of caspase-3.