HMGA2shRNA真核表达载体转染胃癌MKN45细胞株的沉默作用

摘要

Objective The HMGA2shRNA expression vector was transfected into gastric cancer cells MKN45 , to detect the HMGA2mRNA and protein expression. Methods Gastric cancer cells were divided into three groups:blank control group (untransfected cells) .experimental group (cells transfected with HMGA2shRNA) and negative group (cells transfected with missense sequence). The mRNA and protein expression were detected by RT-PCR and Western blot,respectively. Results Compared with the blank group and negative control group,The expression levels of HMGA2mRNA and protein in the experimental group significantly decreased (P < 0.05). Conclusion HMGA2shRNA can suppress the expression of HMGA2. HMGA2shRNA transfected may be used for gastric cancer gene therapy and lay the foundation for the further research.%目的:将HMGA2shRNA真核表达载体转染至胃癌细胞MKN45,检测其HMGA2mRNA及蛋白的表达.方法:实验分3组:空白组(未转染的胃癌细胞)、实验组(转染HMGA2shRNA组)、阴性对照组(转染错义序列组),RT-PCR和Western blot分别检测HMGA2在mRNA及蛋白水平的变化.结果:与空白组和阴性对照组相比,实验组HMGA2mRNA的表达明显下降,差异具有统计学意义(均P＜0.05)；HMGA2蛋白表达的变化与基因的变化相一致,实验组的蛋白表达明显受抑制,差异具有统计学意义(均P＜ 0.05).结论:HMGA2的特异性shRNA表达载体抑制了HMGA2的表达,HMGA2shRNA转染有望用于胃癌的基因治疗,为进一步研究其更多功能打下基础.