微小RNA-451对LN229胶质瘤细胞系侵袭和迁移的影响  

Micro RNA-451 down-regulates glucose transporter-1 to affect LN229 glioma cells invasion and migration

作　　者：郭红宝 南阳[1] 甄英伟[1] 王乐[1] 张亚辉[1] 周西广 周俊虎[1] 韩磊[1] 黄强[1] 俞凯[1] 钟跃[1] 

机构地区：[1]天津医科大学总医院神经外科天津市神经病学研究所神经肿瘤实验室,天津300052

出　　处：《中华神经医学杂志》2016年第6期548-553,共6页Chinese Journal of Neuromedicine

基　　金：天津市应用基础与前沿技术研究计划（13JCQNJC10800、13JCZDJC31000）

摘　　要：目的研究微小RNA-451（miRNA-451）下调葡萄糖转运蛋白1（GLUT1）对人脑LN229胶质瘤细胞系侵袭和迁移的影响。方法体外常规培养LN229细胞，将其分为空白对照组、无义序列组及miRNA-451组，空白对照组不进行任何处理，后2组分别转染无义序列和miRNA-451拟似物序列。转染48h后，RT-PCR检测3组细胞miRNA-451的表达；Western blotting检测转染后细胞腺苷酸活化性蛋白激酶（AMPK）、GLUT1蛋白的表达：细胞免疫荧光染色检测转染后细胞膜蛋白GLUT1的表达：划痕实验检测细胞的迁移能力：Transwell实验检测细胞的侵袭能力。结果与空白对照组和无义序列组相比，miRNA-451组细胞miRNA-451mRNA的表达量明显增高，AMPK和GLUT1蛋白的表达明显减低，膜蛋白GLUT1荧光强度明显减弱，划痕修复率和穿膜细胞数（126．23±4．52、120.31±3．61VS48．42±2.31）明显降低，差异有统计学意义（P〈0．05）。结论miRNA-451可能通过调控AMPK下调GLUT1抑制人脑胶质瘤细胞的侵袭和迁移能力。Objective To investigate the influence of glucose transporter-1 （GLUT-1） down-regulation by micro RNA-451 （miRNA-451） in invasion and migration of LN229 glioma cells. Methods The human glioblastoma cell line LN229 was routinely cultured in vitro; and they were divided into blank control group, scramble group and miRNA-451 group; cells in the blank control group did not give any treatment, and cells in the scramble group and miRNA-451 group were given nonsense sequence or miRNA-451 sequence. The miRNA-451 expression was identified by real time-PCR 48 h after transfection, and AMPK and GLUT1 protein expressions were evaluated by Western blotting. Fluorescence intensity changes of GLUT1 from cell membrane were detected by immunofluorescence. Wound-healing assay was employed to detect the cell migration ability. The cell invasion ability was determined by Transwell assay. Results As compared with blank control group and scramble group, miRNA-451 group had markedly up-regulated miRNA-451 mRNA expression, obviously decreased AMPK and GLUT1 protein expressions, with statistical differences （P〈0.05）. Immunofluorescence indicated that fluorescence intensity of membrane GLUT1 in the miRNA-451 group decreased significantly as compared with that in the blank control group and scramble group （P〈0.05）; Wound-healing assay indicated that cell scratch repair rate obviously reduced and number of cells through Transwell chambers significantly decreased in the miRNA-451 group as compared with those in the other two groups, with statistical significant differences （P〈0.05）. Conclusions MiRNA-451 can inhibit the migration and invasion of glioma cells via AMPK down-regulation of GLUT 1 expression level.

关 键 词：微小RNA-451 神经胶质瘤 葡萄糖转运蛋白1 迁移 侵袭 

分 类 号：R739.41[医药卫生—肿瘤]