摘要
目的探讨微小RNA-199a(miR-199a)调节缺氧诱导因子-1α(HIF-1α)表达对缺氧状态下胃癌细胞上皮间充质转化(EMT)的影响及可能的机制。方法低氧处理胃癌细胞24h,通过转染si-HIF-1α和miR-199amimic,采用实时荧光定量聚合酶链式反应(PCR)检测miR-199a的表达水平,蛋白质印迹法(Western blotting)检测HIF-1α及相关EMT蛋白的变化,镜下观察胃癌细胞形态。结果低氧处理后,能下调miR-199amRNA表达,上调HIF-1α和间叶细胞标志蛋白表达,下调E-钙黏素(E-cadherin)蛋白表达。转染miR-199amimic后能抑制HIF-1α的蛋白表达,转染si-HIF-1α和miR-199amimic后能部分逆转低氧的作用,抑制EMT过程的发展。结论 miR-199a可能通过降低HIF-1α的表达抑制缺氧状态下胃癌细胞的EMT过程的发生。
Objective To investigate the effect of microRNA-199a（miR-199a） on hypoxia epithelial-mesenchymal transition （EMT） of gastric Carcinoma by regulating hypoxia inducible factor-1α（HIF-1α） gene expression and its probable mechanism. Meth- ods After hypoxia for 24 h,si-HIF-1α and miR-199a mimic were transfected to gastric cancer cells. The expression levels of miR- 199 was detected by using quantitative real-time PCR. The levels of HIF-1α and related protein in EMT were examined by Western blotting. Gastric cancer cell morphology were observed with an inverted microscope. Results After hypoxia,the expression levels of miR-199a mRNA and E-cadherin protein were decreased,and the expression levels of HIF-1α and mesenchymal cell marker protein were increased. After transfection of miR-199a mimic, the expression of HIF-1α was inhibited. Then, the transfection of si-HIF-1α and miR-199a mimic could partially reverse the effect of hypoxia and inhibit the development of EMT process. Conclusion miR-199a could suppress the transition process and EMT process under hypoxia,it is may be achieved by down-regulating HIF-1α gene expression.
出处
《重庆医学》
CAS
北大核心
2016年第35期4914-4916,4921,共4页
Chongqing medicine
