GRASP65的表达对胃癌细胞增殖、凋亡、迁移和侵袭的影响  

Effect of GRASP65 expression on proliferation,apoptosis,migration and invasion of gastric cancer cells

作　　者：易永芬[1] 张婉玉[1] 彭豆豆 罗祎[2] 

机构地区：[1]重庆医科大学基础医学院病理学教研室分子医学与肿瘤研究中心,重庆400016 [2]重庆医科大学附属第一医院妇产科,重庆400016

出　　处：《重庆医科大学学报》2017年第12期1614-1619,共6页Journal of Chongqing Medical University

摘　　要：目的:探讨高尔基体堆叠蛋白65(Golgi reassembly stacking protein 65,GRASP65)在体外对人胃癌细胞增殖、凋亡、迁移和侵袭能力的影响。方法:首先,通过Western blot和RT-qRCR鉴别低分化胃癌细胞MKN-45、中分化胃癌细胞SGC-7901、高分化胃癌细胞MKN-28和正常胃黏膜细胞GES-1中GRASP65蛋白和mRNA的表达量。然后,用si RNA干扰技术通过Lipofectamine 2000瞬时转染下调和MKN-45细胞中GRASP65的表达量。用qRT-PCR和Western blot检测转染后各组细胞中GRASP65的蛋白和mRNA表达量以证实转染成功。最后,用CCK-8、流式细胞术、Transwell迁移和侵袭实验检测转染后的各组胃癌细胞增殖、凋亡、迁移和侵袭能力的改变。结果:Western blot和qRT-RCR结果显示MKN-45细胞和SGC-7901细胞中GRASP65的蛋白和mRNA表达量明显高于MKN-28细胞和GES-1细胞中GRASP65蛋白和mRNA表达量(P<0.05),故选择低分化胃癌细胞MKN-45作为实验细胞株来下调GRASP65的表达量。qRT-PCR和Western blot结果证明转染成功,转染组GRASP65的mRNA和蛋白表达量与阴性对照组和空白对照组对比有统计学意义(P<0.05)。CCK-8、流式细胞术、Transwell迁移和侵袭实验均证明,下调MKN-45细胞的GRASP65的表达量可抑制胃癌细胞MKN-45体外增殖、迁移和侵袭能力,并促进细胞凋亡。结论:下调GRASP65的表达可明显减弱胃癌细胞的体外增殖能力,同时促进胃癌细胞的凋亡、体外侵袭和迁移。Objective：To investigate the effect of GRASP65 expression on proliferation,apoptosis,migration and invasion of gastric cancer cells in vitro.Methods：First,Western blot and qRT-PCR were carried out in three different differentiated gastric cancer cells（MKN-45,SGC-7901,MKN-28）and normal gastric mucosa cells（GES-1）to test the expression of GRASP65 protein and mRNA.Then,si RNA interference technique was used through Lipofectamine 2000 to transient transfection MKN-45 cells to knockdown GRAPS65 expression.Western blot and qRT-PCR were used to test GRAPS65 expression and confirm transfection success.Finally,CCK-8,flow cytometry,Transwell migration and invasion experiments were used to test the change of ability of cell proliferation,apoptosis,migration and invasion after transfection.Results：Western blot and qRT-PCR results showed that the expressions of GRASP65 protein and mRNA in MKN-45 and SGC-7901 were much higher than those in MKN-28 and GES-1.MKN-45 was chosen as experimental cells.Western blot and qRT-PCR results demonstrated that GRASP65 had been efficiently downregulated through si RNA tansfection.In addition,CCK-8,flow cytometry,Transwell chamber assays results showed that,GRASP65 knockdown can suppress growth,promote apoptosis,inhibit migration and invasion of gastric cancer cells in vitro.Conclusion：GRASP65 knockdown could significantly suppress growth,promote apoptosis,inhibit migration and invasion of gastric cancer cells MKN-45 in vitro.

关 键 词：GRASP65 SIRNA 增殖 凋亡 迁移 侵袭 

分 类 号：R735.2[医药卫生—肿瘤]