抑制高迁移率族蛋白A2对肾癌细胞侵袭转移能力的影响

摘要

目的 观察小干扰 RNA(siRNA)抑制高迁移率族蛋白 A2(HMGA2)基因表达对人肾癌细胞系786-O 迁移、侵袭能力的影响.方法 人肾癌细胞系786-O 扩大培养后分为3组:空白对照组(Ctrl组)、阴性对照组(NC 组)和 siHMGA2组.NC 组、siHMGA2组分别转染阴性对照siRNA和 HMGA2 siRNA,Ctrl组常规培养.采用反转录聚合酶链反应和 Western 免疫印迹法检测转染后 HMGA2 mRNA和蛋白的表达;Western免疫印迹法检测上皮-间质转化(EMT)相关蛋白钙粘蛋白 E和波形蛋白的表达;划痕实验检测细胞迁移能力的变化;Transwell 侵袭实验检测细胞侵袭能力的变化.结果 siHMGA2组 HMGA2 mRNA和蛋白表达水平较 Ctrl 组和 NC组显著下降,差异有统计学意义(P<0.05);siHMGA2组钙粘蛋白 E表达水平较 Ctrl 组、NC组明显升高,波形蛋白表达水平较后两组明显降低,差异有统计学意义(P<0.05);siHMGA2组细胞迁移能力较Ctrl组显著降低,siHMGA2组细胞侵袭能力较 Ctrl 组及 NC组显著降低.结论 抑制 HMGA2的表达可抑制肾癌细胞系786-O 细胞的迁移和侵袭能力,其作用可能是通过抑制 EMT实现的.%Objective To investigate the effect of high mobility group A2 (HMGA2)on cell migration and invasion in renal cell carcinoma. Methods Human renal cell carcinoma cell line 786-O was employed.Cells were divided into three groups:blank group,transfection with negative con-trol siRNA as negative control group,transfection with small interfering RNA of HMGA2 as siHM-GA2 group.HMGA2 mRNA and protein levels were detected by reverse transcription polymerase chain reaction (RT-PCR)and Western blot.The expression of EMT (epithelial-mesenchymal transi-tion)related gene E-Cadherin and Vimentin were detected by Western blotting.Cell scratch assay was performed to examine migration of 786-O cells and Transwell invasion assay was performed to examine invasive capacity of 786-O cells. Results The expression of HMGA2 mRNA and protein were significantly down regulated in siHMGA2 group,compared with the blank group and NC group.The expression of E-cadherin increased while Vimentin decreased significantly in siHMGA2 group compared with other two groups.Cell migratory and invasive capacities of 786-O cells were suppressed significantly in siHMGA2 group compared with other groups. Conclusions Knock-down of HMGA2 gene can effectively inhibit expression of HMGA2 mRNA and protein in 786-O cells,and decrease both cell migratory and invasive capacities as well,probably via the EMT path-way.