敲低锌指E盒增强子结合蛋白1(ZEB1)抑制胃癌细胞的增殖、侵袭和迁移

Knock-down of ZEB1 inhibits the proliferation,invasion and migration of gastric cancer cells

目的通过RNA靶向干扰降低人胃癌BGC823细胞中锌指E盒增强子结合蛋白1(ZEB1)基因表达,观察ZEB1低表达对胃癌BGC823细胞的侵袭、迁移及增殖能力的影响,并检测相关基因lncRNA HOTAIR、上皮钙黏素(E-cadherin)表达情况。方法用载体构建针对人ZEB1基因表达的干扰质粒sh ZEB1,脂质体转染胃癌BGC823细胞后,经G418及有限稀释法筛选稳定转染细胞株。通过实时定量PCR和Western blot法检测BGC823细胞ZEB1 mRNA和蛋白水平,MTT法检测其增殖能力,TranswellTM小室侵袭试验检测转染细胞侵袭能力、划痕试验检测细胞迁移能力,实时定量PCR检测lncRNA HOTAIR、E-cadherinmRNA水平。结果成功构建干扰质粒sh ZEB1,敲低BGC823细胞ZEB1水平后,BGC823细胞的增殖、侵袭和迁移能力降低、lncRNA HOTAIR水平降低、而E-cadherin表达增高。结论靶向降低BGC823胃癌细胞ZEB1的水平,可降低lncRNA HOTAIR水平及细胞增殖、侵袭、迁移力。

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 sh ZEB1 was constructed and transfected into the gastric cancer BGC823 cells by LipofectamineTM2000,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 TranswellTMinvasion 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 sh ZEB1 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.