OIP5-AS1和miR-410对胶质瘤细胞增殖、侵袭及凋亡的影响

Effects of OIP5-AS1 and miR-410 on the proliferation,invasion and apoptosis of glioma cells in vitro

目的通过建立OIP5-AS1 siRNA和miR-410抑制剂转染的U87细胞系,探讨OIP5-AS1和miR-410抑制剂对胶质瘤细胞增殖、侵袭、迁移、凋亡等生物学行为的影响。方法通过Lipofectamine-2000转染人胶质细胞瘤细胞系U87细胞。将细胞分为5组:对照组(未转染细胞)、NC组(转染NC序列的细胞)、OIP5-AS1 siRNA组(转染OIP5-AS1 siRNA的细胞)、miR-410抑制剂组(转染miR-410抑制剂的细胞)、OIP5-AS1 siRNA+miR-410抑制剂(siRNA+inhibitor)组(转染OIP5-AS1 siRNA和miR-410抑制剂的细胞)。通过MTT法检测5组U87细胞的增殖能力,流式细胞术检测细胞周期变化,Annexin V-FITC/PI双染色法检测细胞凋亡,Transwell实验检测细胞侵袭力,Wound-healing实验检测细胞迁移。结果OIP5-AS1 siRNA组细胞增殖较对照组明显受到抑制(P<0.05)。miR-410抑制剂组细胞增殖较对照组明显增强(P<0.05)。与OIP5-AS1 siRNA组比较,siRNA+inhibitor组增殖显著增加(P<0.05)。与对照组比较,OIP5-AS1 siRNA组G0/G1期的细胞比例显著增高,而S期的细胞比例降低,细胞凋亡率明显升高差异均有统计学意义(P<0.05);在miR-410抑制剂组G0/G1期的细胞比例降低,S期的细胞比例显著升高,细胞凋亡率明显降低差异均有统计学意义(P<0.05)。与OIP5-AS1 siRNA组比较,siRNA+inhibitor组G0/G1期细胞明显减少,S期细胞比例增加,凋亡率降低(P<0.05)。各组G2期所占比例无显著差异。OIP5-AS1 siRNA组的侵袭细胞数量和划痕修复能力显著降低,而miR-410抑制剂组的侵袭细胞数量和划痕修复能力显著升高(P<0.05)。siRNA+inhibitor组在这两个指标上均高于OIP5-AS1 siRNA组(P<0.05)。结论沉默OIP5-AS1可抑制U87细胞的增殖、侵袭和迁移。下调OIP5-AS1表达可诱导U87细胞在G0/G1期停滞和凋亡。miR-410抑制剂可逆转OIP5-AS1 siRNA对U87细胞的生物学作用。

Objective To investigate the effects of OIP5-AS1 and miR-410 inhibitor on the proliferation,invasion,migration and apoptosis of glioma cells by establishing the U87 cell line transfected with OIP5-AS1 siRNA and miR-410 inhibitor.Methods Lipofectamine 2000 was used to transfect human glioma cell line U87 cells.The cells were divided into 5 groups:control group(untransfected cells),NC group(cells transfected with NC sequence),OIP5-AS1 siRNA group(cells transfected with OIP5-AS1 siRNA),miR-410 inhibitor group(cells transfected with miR-410 inhibitor),OIP5-AS1 siRNA+miR-410 inhibitor(siRNA+inhibitor)group(cells transfected with OIP5-AS1 siRNA and miR-410 inhibitor).The proliferation ability of U87 cells in the five groups was detected by MTT method,and cell cycle changes were detected by flow cytometry.Apoptosis was detected by Annexin V-FITC/PI double staining method,cell invasiveness was detected by Transwell experiment,and cell migration was detected by Wound-healing experiment.Results The cell proliferation of the OIP5-AS1 siRNA group was significantly inhibited compared with the control group(P<0.05).The cell proliferation in the miR-410 inhibitor group was significantly higher than that in the control group(P<0.05).Compared with the OIP5-AS1 siRNA group,the proliferation of the siRNA+inhibitor group was increased significantly(P<0.05).Compared with the control group,the proportion of cells in the G0/G1 phase of the OIP5-AS1 siRNA group was significantly increased,while the proportion of cells in the S phase was reduced,and the apoptosis rate was significantly increased(P<0.05).The proportion of cells in the G0/G1 phase of the miR-410 inhibitor group was decreased,the proportion of cells in the S phase was significantly increased,and the apoptosis rate was significantly reduced(P<0.05).Compared with the OIP5-AS1 siRNA group,cells in the G0/G1 phase of the siRNA+inhibitor group were significantly reduced,the proportion of cells in the S phase was increased,and the apoptosis rate was reduced(P<0.05).There was no significant difference in the proportion of cells in the G2 phase in each group.The number of invasion cells and scratch repair ability in the OIP5-AS1 siRNA group were significantly reduced,while those in the miR-410 inhibitor group were significantly increased(P<0.05).The above two indexes in siRNA+inhibitor group were higher than those in the OIP5-AS1 siRNA group(P<0.05).Conclusion Silencing OIP5-AS1 can inhibit the proliferation,invasion and migration of U87 cells.Down-regulating the expression of OIP5-AS1 can induce U87 cell apoptosis and G0/G1 phase cell cycle arrest.miR-410 inhibitor can reverse the biological effects of OIP5-AS1 siRNA on U87 cells.