二甲双胍对人胰腺癌细胞株PANC-1生长侵袭影响及其机制的探讨  

Effects of metformin on proliferation and invasion of human pancreatic cell line PANC-1 and mechanisms

作　　者：唐曦平[1] 唐国都[1] 方春芸[1] 张露艺[1] 梁志海[1] 

机构地区：[1]广西医科大学第一附属医院消化内科,广西南宁530021

出　　处：《中华肿瘤防治杂志》2013年第13期963-967,共5页Chinese Journal of Cancer Prevention and Treatment

基　　金：国家自然科学基金(81060043);广西卫生厅课题(Z2012104);广西教育厅课题(201204LX048);广西2011年研究生教育创新计划(2011105981002D31)

摘　　要：目的:研究二甲双胍对人胰腺癌细胞株PANC-1增殖、凋亡及侵袭的影响,并初步探讨其可能机制。方法:体外培养人胰腺癌细胞株PANC-1,给予不同浓度二甲双胍进行干预作为实验组,无药物组作为对照组。MTT法检测二甲双胍对PANC-1细胞存活率的影响;流式细胞仪检测二甲双胍对PANC-1细胞周期及细胞凋亡的影响;Transwell小室侵袭实验观察细胞侵袭能力;RT-PCR检测Bax、Bcl-2、Caspase-3、Cyclin D1、MMP-2和MMP-9mRNA的表达。结果:与对照组相比,用药组细胞增殖活性明显下降,F值分别为70.318、327.201和654.196,P<0.01,且呈时间-浓度依赖性。流式细胞仪检测细胞周期结果显示,随着二甲双胍浓度的增加,G0/G1期细胞所占百分率逐渐增加,S期和G2/M期细胞所占百分率逐渐下降,F值分别为208.365、195.308和48.87,P<0.01;流式细胞仪分析对照组与实验组早期和晚期细胞凋亡率差异无统计学意义,F值分别为0.123和0.298,P>0.05。侵袭实验结果显示,随着二甲双胍浓度的增加,细胞侵袭能力逐渐下降,F=66.131,P<0.01。RT-PCR示,二甲双胍干预组Cyclin D1、MMP-2和MMP-9mRNA表达明显降低,t值分别为6.789、13.452和25.72,P<0.01;Bax、Bcl-2和Caspase-3mRNA与对照组相比差异无统计学意义,t值分别为-0.683、-1.567和0.78,P>0.05。结论:二甲双胍能显著抑制人胰腺癌细胞株PANC-1的增殖和侵袭,机制主要与其阻滞细胞周期以及影响相关基因表达有关;二甲双胍对PANC-1细胞的凋亡无明显诱导作用。OBJECTIVE： To investigate the effects of metformin on proliferation,apoptosis and invasion in human Pancreatic Cell Line PANO1, and appraise the possible potential mechanism. METHODS： Human pancreatic cell line PANC-1 were Cultured in vitro ,and were treated with various concentration of metformin as experimental group or with- out metformin as control group. Survival rate of PANC-lcells was determined by Methyl thiazolyl tetrazolium （MTT） as- say,and the cell cycle changes were analyzed by flow cytometry. Cell apoptosis was determined by flow cytometry. Cells invasion viality was measured by transwell chamber experiment. Expressions of Bax, Bcl-2, Caspase-3, CyclinD1, MMP-2, MMP-9 mRNA were deteced by RT-PCR. RESULTS：Compared with the control,metformin decreased the proliferation of PANC-1 cells in a dose- and time-dependent manner（F were 70. 318,327. 201,654. 196,P〈0.01）. With the increase of the concentration of metformin,the percentage of cells in Go/G1 phase increased gradually, and the percentage of cells in S and G2/M decreased gradually （F were 208. 365,195. 308,48.87, P〈0.01）. There is no significant difference of early or late apoptosis rate in the control group and the experimental group（F were 0. 123,0. 298, P〉0.05）. With the increase of the concentration of metformin, the invasion viality of PANC-1 cells decreased gradually（F = 66.131, P〈 0.01 ）. The ex- pression of CyclinD1 ,MMP-2 ,MMP-9 mRNA was down-regulated（t were 6. 789,13. 452,25.72, P〈0.01）. The expres- sion of Bax, Bcl-2 and Caspase-3 mRNA was not altered significantly（t were - 0. 683, - 1. 567,0.78, P〉 0.05）. CON-

关 键 词：胰腺肿瘤 二甲双胍 人胰腺癌细胞株PANC-1 细胞增殖 细胞凋亡 肿瘤浸润 流式细胞术 

分 类 号：R735.9[医药卫生—肿瘤]