E2F-1过表达对人胃癌SGC-7901细胞增殖的影响及其机制  

Effects of E2F-1 overexpression on the proliferation of human gastric cancer cell line SGC-7901 and its mechanism

作　　者：曹稳珑 韦尉元[1] 张笑石 罗文[1] 严林海[1] 谢玉波[2] 肖强[1] 

机构地区：[1]广西医科大学第一附属医院胃肠腺体外科,南宁530021 [2]广西医科大学第一附属医院麻醉科,南宁53002

出　　处：《广东医学》2014年第11期1654-1658,共5页Guangdong Medical Journal

基　　金：国家自然科学基金资助项目(编号:30860273);广西科学研究与技术开发计划项目(编号:桂科攻10124001A-22);广西自然科学基金资助项目(编号:2013GXNSFAA019163)

摘　　要：目的探讨E2F-1过表达对胃癌SGC-7901细胞增殖、生长和细胞周期的影响及其分子机制。方法采用携带E2F-1基因的重组慢病毒颗粒(LV-E2F-1-GFP)感染人胃癌SGC-7901细胞(LV-E2F-1-GFP组);以对照慢病毒颗粒(LV-GFP)感染人胃癌SGC-7901细胞作为阴性对照(LV-GFP组);空白对照组常规培养胃癌SGC-7901细胞,不作任何处理。分别采用CCK-8法检测细胞的增殖活力,流式细胞仪检测各组细胞周期的分布,RT-PCR和Western blot技术检测细胞中Skp2、Bax、Bcl-2、CylinD1和Survivin基因mRNA和蛋白的表达。结果与LV-GFP组和空白对照组比较,LV-E2F-1-GFP组人胃癌SGC-7901细胞增殖活力明显降低(P<0.05),G0/G1期所占比例上升(P<0.05),Skp2、Bcl-2、CylinD1和Survivin基因的mRNA和蛋白的表达降低(P<0.05),Bax基因的mRNA和蛋白的表达上调(P<0.05),而LV-GFP组和空白对照组比较,差异无统计学意义(P>0.05)。结论慢病毒介导E2F-1基因过表达抑制胃癌细胞增殖和生长,使细胞周期停滞在G0/G1期,其机制可能与E2F-1基因过表达使胃癌细胞Skp2、Survivin、Bcl-2、CylinD1基因表达下调和Bax基因表达上调有关。Objective To investigate the effect of E2F -1 overexpression on the proliferation,growth and cell cy-cle of human gastric cancer cell line SGC -7901 and the molecular mechanism. Methods Humans gastric cancer SGC -7901 cells were infected by E2F - 1 recombinant lentiviral particles（LV - E2F - 1 - GFP）,as experimental group（LV -E2F - 1 - GFP group）,while control lentiviral particles（LV - GFP）infected human gastric cancer SGC - 7901 cells,as a negative control group（LV - GFP group）and blank control group was cultured conventionally without any treatment. CCK - 8 were used to detect cell proliferation activity,while flow cytometry was used for assessment of cell cycle distribu-tion. Semi - quantitative reserve transcription - polymerase chain reaction（RT - PCR）and Western blot were applied for measurement of mRNA and protein expression of Skp2,Bax,Bcl - 2,CylinD1 and Survivin. Results Compared with LV - GFP group and blank control group,significant lower proliferation activity（P 〈 0. 05）,increased G0 / G1 phase pro-portion（P 〈 0. 05）,down - regulation of mRNA and protein of Skp2,Bcl - 2,CylinD1 and Survivin（P 〈 0. 05）,up -regulation of Bax mRNA and protein（P 〈 0. 05）,were revealed LV - E2F - 1 - GFP group. Furthermore,there was no significant difference between LV - GFP group and blank control group（P 〉 0. 05）. Conclusion Lentivirus - mediated overexpression of E2F - 1 induces growth inhibition of cancer cells,and cell cycle arrest at G0 / G1 phase;probably via up - regulation of Bax and down - regulation of Skp2,Survivin,Bcl - 2 and CylinD1.

关 键 词：胃癌 SGC-7901细胞 E2F-1 过表达 

分 类 号：R735.2[医药卫生—肿瘤]