siRNA沉默高迁移率蛋白A2基因对人膀胱移行细胞癌细胞T24增殖凋亡的影响

Effects of HMGA2 gene silencing by small interfering RNA on the proliferation and apoptosis of human bladder cancer cell T24 in vitro

目的 观察siRNA靶向沉默HMGA2基因在人膀胱移行细胞癌T24细胞中表达的变化,探讨抑制HMGA2基因对T24细胞增殖、周期和凋亡的影响.方法 针对人HMGA2基因构建siRNA干扰片段,瞬时转染T24细胞后,采用Western-blot方法检测转染siRNA干扰片段48h后T24细胞蛋白表达量的变化,CCK-8法检测T24细胞增殖和流式细胞仪（FCM）检测T24细胞周期和凋亡情况.结果 转染后48h的T24细胞HMGA2蛋白表达水平较空白对照组（CON）和阴性对照组（NC）显著下降,差异有统计学意义（P＜0.05）,转染HMGA2-siRNA的实验组抑制T24细胞增殖,T24细胞S期细胞比例（35.47±0.23）％高于CON组和NC组（P＜0.05）,实验组T24细胞凋亡率为（17.25±0.24）％,明显高于CON组和NC组（P＜0.05）.结论 HMGA2基因的特异性siRNA可以抑制T24细胞增殖,细胞周期阻滞在S期,并促进其凋亡.

Objectives To investigate the effects of HMGA2 gene silencing by small interfering RNA on the proliferation,cell cycle and apoptosis of human bladder cancer cell T24 invitro.Methods Using LipofectamineTM2000,HMGA2 siRNA was transfected into T24 cells,Western blot was used to detect the expression of HMGA2 protein.Cell proliferation activity was analyzed using cell counting kit-8（CCK-8）,The cell cycle and apoptosis rate of T24 cells were determined by using flow cytometry assays.Results The siRNA silencing effect was observed by treatment of siRNA HMGA2.After HMGA2-siRNA transfection,the protein expression of HMGA2 in experiment group was significantly lower than that of CON and NC group.The growth rate of T24 cells treated by siRNA was much slower than the CON and NC group,the difference was satistically significant.The percentage of S phase cells was sigificantly increased（P ＜ 0.05）.The apoptasis rate of experimental group was（17.25 ± 0.24） ％,which was signicantly increased than those of CON and NC group（P ＜0.05）.Conclusions siRNA-mediated HM-GA2 gene silencing can efectively inhabit proliferation of T24 cells,induced the accumulated of T24 cells in the S phase and promote their apoptosis.