转染葡萄糖神经酰胺合成酶小干扰RNA的胃癌细胞株SGC7901、SGC7901/DDP凋亡观察  

Observation on apoptosis of gastric cancer cell line SGC7901 and cisplatin-resistant gastric cancer cell line SGC7901/DDP transfected with glucosylceramide synthase siRNA

作　　者：张俊杰[1] 葛艳丽[1] 王声乐 王喆[1] 王志荣[1] ZHANG Junjie;GE Yanli;WANG Shengyue;WANG Zhe;WANG Zhirong(Tongji,Hospital of Tongji University,Shanghai 200065,China)

机构地区：[1]同济大学附属同济医院,上海200065

出　　处：《山东医药》2020年第33期6-9,共4页Shandong Medical Journal

基　　金：上海市卫生与计划生育委员会青年科学基金项目(20124Y100)。

摘　　要：目的观察转染葡萄糖神经酰胺合成酶(GCS)小干扰RNA的胃癌细胞株SGC7901、顺铂耐药胃癌细胞株SGC7901/DDP凋亡情况,并探讨其机制。方法取正常胃黏膜上皮细胞株GES-1(GES-1组)、人胃癌细胞株SGC7901(SGC7901组)、顺铂耐药人胃癌细胞株SGC7901/DDP(SGC7901/DDP组),转染GCS小干扰RNA后继续培养48 h,使用流式细胞仪检测各组凋亡细胞数,计算凋亡指数;采用Real-time PCR法检测各组细胞中MDR-1、BCL2、BAX mRNA,采用Western blotting法检测各组细胞中P-糖蛋白(P-gp)、BCL2、BAX、Caspase3蛋白。结果SGC7901组、SGC7901+SiRNA组细胞凋亡指数分别为0.36±0.05、14.98±1.01,两组相比,P<0.05;SGC7901/DDP组、SGC7901/DDP+SiRNA组细胞凋亡指数分别为0.18±0.02、12.75±0.68,两组相比,P<0.05。SGC7901/DDP+SiRNA组、SGC7901+SiRNA组细胞凋亡指数分别为12.75±0.68、14.98±1.01,两组相比,P<0.05。SGC7901组细胞中MDR-1、BCL2 mRNA的相对表达量高于SGC7901+SiRNA组(P<0.05),BAX mRNA的相对表达量低于SGC7901+SiRNA组(P<0.05);SGC7901/DDP组细胞中MDR-1、BCL2 mRNA的相对表达量高于SGC7901/DDP+SiRNA组(P<0.05),BAX mRNA的相对表达量低于SGC7901/DDP+SiRNA组(P<0.05);SGC7901/DDP组细胞中MDR-1 mRNA的相对表达量高于SGC7901组(P<0.05),SGC7901/DDP+SiRNA组细胞中MDR-1 mRNA的相对表达量高于SGC7901+SiRNA组(P<0.05)。SGC7901组细胞中P-gp、BCL2蛋白的相对表达量高于SGC7901+SiRNA组(P<0.05),BAX、Caspase3蛋白的相对表达量低于SGC7901+SiRNA组(P<0.05);SGC7901/DDP组细胞中P-gp、BCL2蛋白的相对表达量高于SGC7901/DDP+SiRNA组(P<0.05),BAX、Caspase3蛋白的相对表达量低于SGC7901/DDP+SiRNA组(P<0.05);SGC7901/DDP组细胞中P-gp蛋白的相对表达量高于SGC7901组(P<0.05),SGC7901/DDP+SiRNA组细胞中P-gp蛋白的相对表达量高于SGC7901+SiRNA组(P<0.05)。结论转染GCS SiRNA可促进胃癌细胞株SGC7901、胃癌顺铂耐药株SGC7901/DDP凋亡,其机制可能与调节凋亡通路中的BCL2、BAX、Caspase3蛋白表达有关;Objective To observe the apoptosis of gastric cancer cell line SGC7901 and cisplatin-resistant gastric cancer cell line SGC7901/DDP transfected with glucosylceramide synthase(GCS)siRNA,and to explore the mechanisms.Methods Normal gastric epithelial cell line GES-1(GES-1 group),human gastric cancer cell line SGC7901(SGC7901 group),cisplatin-resistant human gastric cancer cell line SGC7901/DDP(SGC7901/DDP group)were transfected with GCS siRNA and cultured for 48 h.The apoptosis rates were evaluated by flow cytomerty.The expression levels of MDR-1,BCL2 and BAX gene were detected by RT-PCR,and the expression levels of P-gp,BCL2,BAX,and Caspase-3 protein were detected by Western blotting.Results The apoptosis indexes of SGC7901 group and SGC7901+SiRNA group were0.36±0.05 and 14.98±1.01,with statistically significant difference(P<0.05).The apoptosis indexes of SGC7901/DDP group and SGC7901/DDP+SiRNA group were 0.18±0.02 and 12.75±0.68,with statistically significant difference(P<0.05).The apoptosis indexes of SGC7901/DDP+SiRNA group and SGC7901+SiRNA group were 12.75±0.68 and14.98±1.01,with statistically significant difference(P<0.05).The relative expression of MDR-1 and BCL2 mRNA in SGC7901 group was higher than that in SGC7901+siRNA group(P<0.05),and the Bax mRNA was lower than that of SGC7901+siRNA group(P<0.05).The relative expression of MDR-1 and BCL2 mRNA in SGC7901/DDP group was higher than that in SGC7901/DDP+siRNA group(P<0.05),and the BAX mRNA was lower than that in SGC7901/DDP+siRNA group(P<0.05).The relative expression of MDR-1 mRNA in SGC7901/DDP group was higher than that in SGC7901 group(P<0.05),and that in SGC7901/DDP+siRNA group was higher than that in SGC7901+siRNA group(P<0.05).The relative expression of P-gp and BCL2 protein in SGC7901 group was higher than that in SGC7901+siRNA group(P<0.05),and the expression of Bax and Caspase-3 protein was lower than that of SGC7901+siRNA group(P<0.05).The relative expression of P-gp and BCL2 protein in the SGC7901/DDP group was higher than that in SGC7901/

关 键 词：葡萄糖神经酰胺合成酶 小干扰RNA 胃癌 肿瘤多药耐药 细胞凋亡 细胞实验 

分 类 号：R735.2[医药卫生—肿瘤]