慢病毒介导的靶向骨桥蛋白的siRNA对人脑胶质瘤细胞侵袭和凋亡的影响

目的 研究骨桥蛋白(osteopontin,OPN)下调后对胶质瘤细胞侵袭和凋亡的影响.方法 在体外以慢病毒介导的OPN小干扰RNA(small interference,siRNA)感染胶质瘤细胞系U251细胞,实时PCR和Western印迹检测OPN表达.通过Transwell实验、流式细胞仪检测,观察OPN表达下调后对胶质瘤细胞侵袭和细胞凋亡的影响.结果 慢病毒介导的OPN siRNA感染能显著降低U251细胞OPNmRNA水平及蛋白表达,有效抑制U251细胞侵袭能力,并诱导其凋亡,OPN siRNA感染组中Bcl-2、尿激酶型纤溶酶原激活剂(uPA)、MMP-2和MMP-9明显减少,Bax显著增多.结论 慢病毒介导的OPNsiRNA可显著抑制U251细胞的侵袭,并促进其凋亡.

Effect of osteopontin silencing by lentivirus-mediated delivery of siRNA on glioma cell invasion and apoptosis

Objective To investigate the effect of osteopontin silencing on the invasion and apoptosis of U251 cells. Methods The invasion, apoptosis and levels of uPA, MMP-2 and MMP-9 were determined by invasion assay, flow cytometry, Western blot and real-time fluorescence quantitative PCR respectively.Results Osteopontin small interference (siRNA) inhibited osteopontin expression and cell invasion,promoted apoptosis in U251 cells. In addition, the expression of Bcl-2, uPA, MMP-2 and MMP-9 was decreased, while Bax level was elevated. Conclusion Osteopontin siRNA can inhibit U251 cells invasion via the down-regulation of uPA, MMP-2 and MMP-9 levels, and promote apoptosis through induction of Bax expression and inhibition of Bcl-2 level. It suggests that osteopontin plays an important role in human glioma progression.