RNA干扰高迁移率族蛋白A2沉默对裸鼠肾癌细胞上皮-间充质转化的影响

Effect of HMGA2 silencing by RNA interference on epithelial-mesenchymal transition of renal carcinoma cells in nude mice

目的:通过动物实验观察沉默高迁移率族蛋白A2(high mobility group A2,HMGA2)表达对人肾癌细胞ACHN成瘤的影响。方法:通过慢病毒转染技术,培养HMGA2沉默的ACHN细胞。实验分三组:空白组、阴性对照组、HMGA2-siRNA组,建立裸鼠移植瘤模型,取瘤称重,比较各组的成瘤体积、质量和成瘤时间。用实时荧光定量聚合酶链反应(qRT-PCR)和蛋白印迹法(Western blot)检测上皮-间充质转化(epithelial-mesenchymal transition,EMT)中E-钙黏蛋白(E-cadherin)、N-钙黏蛋白(N-cadherin)和转录因子(Snail)的mRNA及蛋白表达。结果:成功构建特异性沉默HMGA2表达的肾癌细胞株,选择HMGA2-siRNA2进行后续试验。与空白组和阴性对照组相比,HMGA2-siRNA组成瘤体积和质量明显降低,成瘤时间显著延长,差异具有统计学意义(P<0.001)。RNA干扰沉默HMGA2表达后,E-cadherin的mRNA和蛋白表达均增高,N-cadherin和Snail的mRNA及蛋白表达均降低,差异具有统计学意义(P<0.001)。结论:HMGA2-siRNA能够特异性沉默ACHN细胞中的HMGA2基因,并且HMGA2能够促进ACHN细胞的EMT,进而推动肾癌的发生发展。

Objective:To investigate the influence of silencing high mobility group A2(HMGA2)on the tumorigenesis of human renal carcinoma ACHN cells by observing animal experiment.Methods:HMGA2-silenced ACHN cells were cultured by lentivirus transfection technique.The experiment was divided into three groups,including untreated group,control-siRNA group and HMGA2-siRNA group.The transplanted tumor model of nude mice was established.The tumor was weighed,and the volume,mass and time of tumor formation of each group were compared.The mRNA and protein expression of E-cadherin,N-cadherin and Snail in epithelial-mesenchymal transition(EMT)were detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR)and Western blot.Results:The renal carcinoma cells line specifically silencing the expression of HMGA2 was successfully constructed,and HMGA2-siRNA2 was selected for subsequent tests.The volume and mass of tumor formation in HMGA2-siRNA group were significantly lower than that in untreated group and control-siRNA group,and the time of tumor formation was significantly longer than that in untreated group and control-siRNA group,and the difference was statistically significant(P<0.001).After siRNA inhibited HMGA2 expression,the mRNA and protein levels of E-cadherin increased,and the mRNA and protein levels of N-cadherin and Snail decreased,with statistically significant differences(P<0.001).Conclusion:HMGA2-siRNA can effectively inhibit the expression level of HMGA2 in ACHN cells,and HMGA2 can promote the occurrence of EMT in renal carcinoma cells,thus promoting the occurrence and development of renal carcinoma.