Chen Dengyu, Chu Yifan, Zheng Qingwei, Xu Zhiben, Zhou Ping, Li Sheng
Department of Pathogenic Biology; Anhui Key Laboratory of Infection and Immunity, Bengbu Medical College, Bengbu 233030, China. *Corresponding author, E-mail:
Laboratory Center for Morphology, Bengbu Medical College, Bengbu 233030, China.
Xi Bao Yu Fen Zi Mian Yi Xue Za Zhi. 2017 Aug;33(8):1073-1078.
Objective To down-regulate the expression of zinc-finger E-box binding homeobox 1 (ZEB1) gene by shRNA, and investigate its effect on invasion, migration and proliferation, as well as the related gene expressions of lncRNA HOTAIR and E-cadherin in human gastric cancer BGC823 cells. Methods RNA interfering (RNAi) was used to knock down ZEB1 in gastric cancer BGC823 cells. The recombinant plasmid shZEB1 was constructed and transfected into the gastric cancer BGC823 cells by Lipofectamine 2000, and the stably transfected cells were isolated by G418 selection and limited dilution. The expression of ZEB1 mRNA and protein was detected by real-time quantitative PCR and Western blot analysis. Cell proliferation was determined by MTT assay, and the invasion and migration abilities of BGC823 cells were monitored by Transwell invasion assay and wound healing assay, respectively. The expressions of lncRNA HOTAIR and E-cadherin mRNA were detected by real-time quantitative PCR. Results After ZEB1 expression was successfully down-regulated in BGC823 cells by siRNA, the proliferation, invasion and migration rates in shZEB1 transfection group were significantly lower than those in control group; meanwhile, the expression of lncRNA HOTAIR was reduced and E-cadherin expression was enhanced. Conclusion Knock-down of ZEB1 expression by RNA interference can decease lncRNA HOTAIR expression and restrain cell proliferation, invasion and migration in gastric cancer BGC823 cells.
目的 通过短发夹RNA（shRNA）下调锌指E盒结合同源框1（ZEB1）基因的表达，研究其对人胃癌BGC823细胞侵袭、迁移、增殖以及长链非编码RNA HOTAIR和E-钙黏蛋白相关基因表达的影响。方法 采用RNA干扰（RNAi）技术敲低胃癌BGC823细胞中的ZEB1。构建重组质粒shZEB1，并用Lipofectamine 2000转染至胃癌BGC823细胞，通过G418筛选和有限稀释法分离稳定转染的细胞。采用实时定量PCR和蛋白质免疫印迹分析检测ZEB1 mRNA和蛋白的表达。采用MTT法检测细胞增殖，分别通过Transwell侵袭实验和划痕实验监测BGC823细胞的侵袭和迁移能力。采用实时定量PCR检测lncRNA HOTAIR和E-钙黏蛋白mRNA的表达。结果 通过小干扰RNA（siRNA）成功下调BGC823细胞中ZEB1的表达后，shZEB1转染组的增殖、侵袭和迁移率均显著低于对照组；同时，lncRNA HOTAIR表达降低，E-钙黏蛋白表达增强。结论 RNA干扰敲低ZEB1表达可降低lncRNA HOTAIR表达，抑制胃癌BGC823细胞的增殖、侵袭和迁移。
