Objectives: To investigate effect of MIPU1 silence on proliferation, apoptosis, migration and invasion in U251 cells.
Methods: The shRNA recombinant plasmids targeting MIPU1 gene was transfected into U251 cells. Western blotting was used to identify the inhibitory efficiency at 72 h after transfection. The cell viability was measured by MTT colorimetric assay. Hoechest staining and caspase-3 activity were used to detect apoptosis. Then wound healing assay and transwell migration assay were applied to detect the migration and invasion of cells.
Results: The expression of MIPU1 protein was effectively knocked down in transfected cells (P<0.05). The cellular proliferation was obviously inhibited and apoptosis was increased in shRNA-transfected MIPU1 cells (all P<0.05). The migration and invasion ability of cells transfected with positive plasmid was lower than that in the control group (P<0.05).
Conclusions: Down-regulation of MIPU1 can promote apoptosis while inhibit the proliferation, invasion, and migration of U251 cells.
目的: 探讨特异性短发夹RNA(shRNA)干扰心肌缺血预适应上调基因1(myocardial ischemia preconditioning upregulated gene 1，MIPU1)对人脑星形细胞瘤U251细胞增殖、凋亡、迁移、侵袭能力的影响。方法: 采用瞬时转染MIPU1 shRNA干扰质粒，蛋白质印迹检测其干扰效率，MTT法检测各组细胞的增殖情况，Hoechest染色法和caspase-3活性检测细胞凋亡程度，划痕实验及Transwell实验检测细胞迁移及侵袭能力。结果: 转染MIPU1 shRNA可明显降低MIPU1蛋白的表达(P<0.05)；与对照组比较，MIPU1 shRNA组的U251细胞增殖能力下降，凋亡明显增加，细胞迁移及侵袭能力减弱(均P<0.05)。结论: MIPU1 shRNA能有效降低U251细胞中MIPU1蛋白的表达，干扰MIPU1的表达可促进U251细胞凋亡，抑制U251细胞的增殖、迁移及侵袭。.
Keywords: MIPU1 gene; U251 cell; biological behavior; shRNA interference.