shRNA干扰MIPU1基因对脑星形细胞瘤U251细胞生物学行为的影响  

Effect of MIPU1 gene interference on biological behavior of U251 cell line

作　　者：刘梅冬 王智 孙慧 蒋碧梅 刘可 肖献忠 LIU Meidong;WANG Zhi;SUN Hui;JIANG Bimei;LIU Ke;XIAO Xianzhong(Department of Pathophysiology,Xiangya School of Medcine,Central South University,Changsha 410078,China)

机构地区：[1]中南大学基础医学院病理生理学系

出　　处：《中南大学学报（医学版）》2020年第2期109-114,共6页Journal of Central South University ：Medical Science

基　　金：国家自然科学基金(81770306,81471868)~~

摘　　要：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。Objective: 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).Conclusion: Down-regulation of MIPU1 can promote apoptosis while inhibit the proliferation, invasion, and migration of U251 cells.

关 键 词：MIPU1基因 SHRNA干扰 U251细胞 生物学行为 

分 类 号：R739.41[医药卫生—肿瘤]