靶向Skp2基因siRNA对胶质母细胞瘤U251细胞增殖能力和侵袭性的影响

Effects of targeted Skp2 gene siRNA on the proliferation and invasiveness of glioblastoma U251 cells

目的探讨靶向Skp2基因siRNA对胶质母细胞瘤U251细胞增殖能力和侵袭性的影响。方法 U251分别转染si-SKP2(si-SKP2组)、siNC(siNC组)与PBS(空白组),采用MTT法检测细胞增殖的变化,流式细胞仪检测细胞周期的变化,划痕实验与Transwell侵袭实验检查细胞迁移与侵袭的变化,WB法测定B淋巴细胞瘤-2基因(bcl-2)Bcl-2、基质金属蛋白酶-9(MMP-9)蛋白表达量。结果转染48 h后si-SKP2组、活细胞比例低于siNC组及空白组差异有统计学意义(P<0.05)。相对于空白组与siNC组,si-SKP2组的G2/M期比例显著增加(P<0.05),G0/G1期比例显著降低(P<0.05)。si-SKP2组的48 h迁移距离与过膜细胞数都显著少于空白组和siNC组,差异有统计学意义(P<0.05)。相对于空白组与siNC组,si-SKP2组的Bcl-2、MMP-9蛋白表达量显著降低(P<0.05)。结论靶向Skp2基因siRNA可以抑制胶质母细胞瘤细胞U251的增殖、迁移与侵袭,引起G2/M期阻滞,其作用机制可能与抑制Bcl-2、MMP-9的表达有关。

Objective To investigate the effects of the target Skp2 gene siRNA on the proliferation and invasiveness of glioblastoma U251 cells. Methods The U251 cells were transfected with si-SKP2 (si-SKP2 group),siNC (siNC group) and PBS (blank group),respectively. The changes of cell proliferation were detected by MTT assay,and the changes of cell cycle were measured by flow cytometry,and the changes of cell migration and invasion were measured by scratch test and Transwell,moreover, the expression levels of Bcl-2,MMP-9 protein were detected by WB method. Results At 48 hours after transfection,the proportion of live cells in si-SKP2 group,siNC group and blank group was (75.22±3.19)%,(98.29± 2.11 )% and (100±0)%,respectively,there was significant difference among the three groups ( P <0.05).As compared with that in blank group and the siNC group,the proportion of G2/M phase in si-SKP2 group was increased significantly ( P < 0.05 ),however, the proportion of G0/G1 stage was decreased significantly ( P <0.05).The 48h migration distance and the number of membrane cells in si-SKP2 group were significantly less than those in blank group and siNC group ( P <0.05).As compared with those in blank group and the siNC group,the expression levels of Bcl-2 and MMP-9 protein in si-SKP2 group were decreased significantly ( P <0.05). Conclusion Targeting Skp2 gene siRNA can inhibit the proliferation,migration and invasion of glioblastoma cell U251,and can induce G2/M phase blocking,which may be related to the inhibition of Bcl-2 and MMP-9 expression.