论文部分内容阅读
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 investigate the effect of ZEB1 overexpression on the invasion, migration and proliferation of gastric cancer cell line BGC823 by RNA interference and reduce the expression of ZEB1 gene in human gastric cancer cell line BGC823, and to detect the expression of lncRNA HOTAIR, E-cadherin expression. Methods The plasmids sh ZEB1 targeting human ZEB1 gene were constructed and transfected into BGC823 gastric cancer cells. The stable transfected cell lines were screened by G418 and limited dilution. Real-time quantitative PCR and Western blot were used to detect the mRNA and protein levels of ZEB1 in BGC823 cells. The proliferation of BGC823 cells was detected by MTT assay. The invasion ability of transfected cells was detected by TranswellTM cell invasion assay. Cell migration was assayed by scratch assay. LncRNA HOTAIR, E-cadherin mRNA levels. RESULTS: The knockdown of sh-ZEB1 could inhibit the proliferation, invasion and migration of BGC823 cells, reduce the level of lncRNA HOTAIR, and increase the expression of E-cadherin in BGC823 cells. Conclusion Targeting to reduce the level of ZEB1 in BGC823 gastric cancer cells can reduce the level of lncRNA HOTAIR and cell proliferation, invasion and migration.