摘要
目的:探讨"miRNA海绵"在胃癌细胞系SGC7901上皮-间质转化(EMT)过程中的作用及机制。方法:将人工合成的ZEB2 3'UTR质粒及siZEB2采用脂质体Lipofectamine 2000转染SGC7901细胞。qRT-PCR检测转染后miR-200a/b/c的表达;Tran-swell侵袭实验、划痕实验和克隆形成实验检测细胞侵袭迁移增殖能力;Western Blot检测目的蛋白的表达。结果:与对照组相比,ZEB2 3'UTR转染组miR-200a/b/c的表达均下调,迁移、侵袭、增殖活性均增强;而转染siZEB2后miR-200a/b/c表达均升高,迁移、侵袭、增殖能力则下降。Western Blot显示ZEB2 3'UTR转染导致ZEB2表达升高,E-cadherin表达下降,而Vimentin表达上调;而转染siZEB2后,各项指标则表现出相反的表达趋势。结论:ZEB2 3'UTR可以通过调控miR-200a/b/c的表达调控胃癌细胞EMT过程,调节肿瘤的侵袭与转移。
Objective：To explore the effect and mechanism of miRNA sponge on the epithelial-mesenchymal transition （EMT） of gastric carcinoma cell lines SGC7901. Methods：Synthetic ZEB2 3＇UTR plasmid and siRNA targeting ZEB2 were transfected into the SGC7901 cell line by Lipofectamine 2000. Real-time quantitative polymerase chain reaction was performed to evaluate the expression levels of miR-200a/b/c. Finally, the migratory, invasive, and proliferative activities of the gastric carcinoma cells in vitro were analyzed by the scratch test, the Transwell cell invasion, and the cell cloning assay. The expression of the target protein was detected by Western blot. Results：Compared with the control group, the expressions of miR-200a/b/c significantly decreased, and their migration, invasion, and proliferation capabilities were considerably higher after they were transfected with ZEB2 3＇UTR. Although the expressions of miR-200a/b/c significantly increased, the migratory, invasive, and proliferative activities of SGC7901 cells also degraded after they were transfected with siRNA targeting ZEB2. The expression of ZEB2 increased, and that of E-cadherin decreased at the protein level after they were transfected with ZEB2 3＇UTR. The protein expression of Vimentin in SGC7901 cells significantly increased. The indicators show the opposite trend when cells were transfected with siZEB2, and the differences between the control and mutation groups were insignificant. Conclusion：ZEB2 3＇UTR can regulate EMT course by regulating the miR-200a/b/c expression in gastric carcinoma, consequently regulating the invasion and migration of carcinoma cells.
出处
《中国肿瘤临床》
CAS
CSCD
北大核心
2014年第11期684-688,共5页
Chinese Journal of Clinical Oncology
基金
国家自然科学基金项目(编号:81172356)
天津市基础重点基础科技项目基金(编号:10JCZDJC18500)资助~~
关键词
miRNA海绵
胃癌
微RNAS
上皮-间质转化
侵袭
miRNA sponge, gastric carcinoma, microRNAs, epithelial-mesenchymal transition, invasion
