反义miR-21通过调控hTERT表达抑制胶质瘤细胞生长  

Knockdown of miR-21 inhibits GBM cell growth through regulating hTERT expression

作　　者：李民[1] 李东儒[1] 孙关[2,3] 万政强[2,3] 伏林山[2,3] 

机构地区：[1]南京医科大学附属南京江宁医院神经外科,2111000 [2]南通大学第四附属医院 [3]盐城市第一人民医院神经外科

出　　处：《临床神经外科杂志》2013年第5期277-280,共4页Journal of Clinical Neurosurgery

摘　　要：目的探讨敲低miR-21表达对人胶质瘤细胞系U87细胞功能的影响以及相关作用机制。方法脂质体介导转染miR-21反义寡聚核苷酸(miR-21 inhibitor)敲低U87细胞miR-21的表达。使用实时荧光定量PCR鉴定转染后U87细胞miR-21表达水平;MTT法检测转染后细胞增殖水平,流式法评价转染后细胞周期分布及凋亡变化,并结合Western印迹及RT-PCR验证在U87细胞中miR-21和hTERT间关系。结果体外转染反义miR-21寡聚核苷酸能明显抑制U87细胞生长,诱导其凋亡,并且能够明显下调hTERT表达。结论反义miR-21可能通过下调hTERT表达抑制胶质细胞生长,miR-21可作为胶质瘤基因治疗的靶点。Objective To study the effects of U87 human glioma cell lines growth induced by knockdown of miR-21 in vitro and the possible mechanism. Methods Antisense oligonucleotides of miR-21（ miR-21 inhibitor） were transfected into U87 cells in vitro to knockdown the miR-21expression. Real-time PCR was used to detect the miRNA expression of miR-21 among different treated groups. The proliferation ability was examined by MTT assay between pre and posttransfected cells. Flow cytometry was conducted to discover the cells cycle and apoptosis in different groups. Then,the relationship between miR-21 and hTERT was identified by bioinformatics,Western bolt and RT-PCR assay. Results The expression level of miR-21 was significantly decreased after miR-21 inhibitor treatment. Reduction of miR-21 inhibited cell proliferation,arrested cell cycle,and induced cell apoptosis in U87 GBM cells,with a dramatic decreased expression of human telomerase reverse transcriptase（ hTERT）. Conclusions MiR-21 inhibitor cell growth through regulating hTERT expression. Mi-21 could be used as a therapeutic strategy for GBM treatment and warrants further investigation.

关 键 词：MIR-21 反义寡聚核苷酸 胶质瘤细胞 hTERT 

分 类 号：R739.41[医药卫生—肿瘤]