HMGA2shRNA真核表达载体转染胃癌MKN45细胞株的沉默作用

Short hairpin RNA-mediated silencing of HMGA2 gene in gastric cancer MKN45 cells

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。

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.