shRNA干扰MIPU1基因对脑星形细胞瘤U251细胞生物学行为的影响

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 ischemiapreconditioning 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MIPU1shRNA 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。

Protective effects of Baicalin, Mannitol and Dexamethasone on infective brain edema in rats

Objective: To investigate effect of MIPU1 silence on proliferation, apoptosis, migration andinvasion in U251 cells.Methods: The shRNA recombinant plasmids targeting MIPU1 gene was transfected intoU251 cells. Western blotting was used to identify the inhibitory efficiency at 72 h aftertransfection. The cell viability was measured by MTT colorimetric assay. Hoechest stainingand caspase-3 activity were used to detect apoptosis. Then wound healing assay andtranswell migration assay were applied to detect the migration and invasion of cells.Results: The expression of MIPU1 protein was effectively knocked down in transfectedcells (P<0.05). The cellular proliferation was obviously inhibited and apoptosis wasincreased in shRNA-transfected MIPU1 cells (all P<0.05). The migration and invasion abilityof cells transfected with positive plasmid was lower than that in the control group (P<0.05).Conclusion: Down-regulation of MIPU1 can promote apoptosis while inhibit theproliferation, invasion, and migration of U251 cells.