GRASP65的表达对胃癌细胞增殖、凋亡、迁移和侵袭的影响

发布时间：2018-04-12 13:05

本文选题：GRASP + siRNA　；参考：《重庆医科大学学报》2017年12期

【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(P0.05),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(P0.05)。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。
[Abstract]:Aim: to investigate the effects of Golgi body stack protein 65(Golgi reassembly stacking protein 65 on proliferation, apoptosis, migration and invasion of human gastric cancer cells in vitro.Methods: firstly, Western blot and RT-qRCR were used to identify the expression of GRASP65 protein and mRNA in poorly differentiated gastric cancer cell line MKN-45, moderately differentiated gastric cancer cell line SGC-7901, highly differentiated gastric cancer cell line MKN-28 and normal gastric mucosal cell GES-1.Then, si RNA interference technique was used to down-regulate the transient transfection of Lipofectamine 2000 and the expression of GRASP65 in MKN-45 cells.QRT-PCR and Western blot were used to detect the expression of GRASP65 protein and mRNA in the transfected cells.Finally, the changes of proliferation, apoptosis, migration and invasion of gastric cancer cells after transfection were detected by CCK-8, flow cytometry and Transwell migration and invasion assay.Results the results of Western blot and qRT-RCR showed that the expression of GRASP65 protein and mRNA in MKN-45 cells and SGC-7901 cells was significantly higher than that in MKN-28 cells and GES-1 cells. Therefore, the poorly differentiated gastric cancer cell line MKN-45 was selected as the experimental cell line to down-regulate GRASP65.The results of QRT-PCR and Western blot showed that the transfection was successful.The expression of mRNA and protein in GRASP65 in transfection group was significantly higher than that in negative control group and blank control group. The results of flow cytometry showed that the down-regulation of GRASP65 expression in MKN-45 cells could inhibit the proliferation of gastric cancer cell line MKN-45 in vitro.Migration and invasion ability, and promote apoptosis.Conclusion: down-regulation of GRASP65 expression can significantly reduce the proliferation of gastric cancer cells in vitro, and promote the apoptosis, invasion and migration of gastric cancer cells in vitro.
【作者单位】：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;重庆医科大学附属第一医院妇产科;
【分类号】：R735.2

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