靶向沉默Pax6基因表达抑制视网膜母细胞瘤增殖侵袭的机制研究

摘要

目的 探讨抑制Pax6基因表达对视网膜母细胞瘤增殖侵袭的影响及机制.方法 RT-PCR检测视网膜母细胞瘤及瘤旁组织中Pax6基因的mRNA表达;Control组(不经任何处理)、阴性对照组(转染 NC-siRNA)和Pax6沉默组(转染 Pax6-siRNA)转染人视网膜母细胞瘤HXO-RB44,转染48 h后Western blot检测各组细胞中Pax6、ki67、基质金属蛋白酶2(MMP-2)、磷脂酰肌醇3激酶(PI3K)、磷酸化丝氨酸苏氨酸激酶(p-AKT)蛋白表达;CCK8和Transwell小室分别检测细胞的增殖及侵袭能力.结果 视网膜母细胞瘤中Pax6基因的mRNA表达显著高于瘤旁组织(p<0.01);转染Pax6-siRNA后HXO-RB44细胞中Pax6的蛋白表达显著降低(p<0.01);与Control组及NC-siRNA组比较,Pax6-siRNA组细胞存活率及细胞侵袭数显著降低,ki67、MMP-2、PI3K、p-AKT蛋白表达均显著下调,差异有统计学意义(p<0.01).结论 抑制Pax6基因表达可显著降低视网膜母细胞瘤增殖和侵袭能力,其机制与PI3K/AKT信号通路的调控有关.%Objective To investigate the effect and mechanism of inhibiting Pax6 gene expression on proliferation and invasion of retinoblastoma.Methods The mRNA expression of Pax6 gene in retinoblastoma and its adjacent tissues were detected by RT-PCR;blank control group (without any treatment), negative control group (transfected with NC-siRNA) and Pax6 interference group (transfected with Pax6-siRNA) transfected with human retinoblastoma HXO-RB44, Pax6, ki67, MMP-2, PI3K, p-AKT protein expression transfected for 48h were detected by Western blot;CCK8 and Transwell were used to detect cell proliferation and invasion of cells.Results The mRNA expression of Pax6 gene in retinoblastoma was significantly higher than that in the tumor adjacent tissues (p<0.01).The expression of Pax6 protein after transfeced Pax6-siRNA in HXO-RB44 cells decreased significantly (p<0.01).Compared with the control group and NC-siRNA group, the cell survival rate and cell invasion in Pax6-siRNA group significantly decreased, ki67, PI3K, MMP-2, and p-AKT protein expression decreased significantly (p<0.01).Conclusion Inhibition of Pax6 gene expression can significantly reduce the proliferation and invasion of retinoblastoma, and its mechanism is related to the regulation of PI3K/AKT signal pathway.