MALAT1对胶质瘤细胞增殖迁移的作用及其ceRNA机制研究

The effect of MALAT1 on proliferation and migration of glioma cells and its ceRNA mechanism

目的探讨lncRNA MALAT1在胶质瘤细胞增殖侵袭过程的作用及其与miRNA124和CREB的作用机制。方法培养C_6胶质瘤细胞,设计lncRNA MALAT1的siRNA序列和scramble序列,以50nM浓度干预C_6细胞48小时,并设定空白对照组,使用CCK8法检测各组细胞增殖活力;Transwell检测各组细胞侵袭力;QPCR实验检测各组lnc RNA-MALAT1、miRNA124、CREB基因的表达情况;Western Blot实验检测CREB蛋白的表达情况。结果与正常C_6细胞相比,MALAT1 siRNA干预后,C_6细胞活力减弱(P<0.05),侵袭力降低(P<0.05);MALAT1基因表达降低,miRNA124表达升高,同时CREB mRNA表达降低。Western Blot实验结果显示,各组细胞中CREB蛋白表达与mRNA表达一致。结论在胶质瘤细胞中,lnc-MALAT1发挥关键作用,其可通过调控miRNA124的表达,引起CREB基因转录和蛋白表达升高,从而促进C_6胶质瘤细胞增殖。

Objective To investigate the role of lnc RNA MALAT1 in the proliferation and invasion of glioma cells and its mechanism with miRNA124 and CREB. Methods C6 glioma cells were cultured in vivo,The MALAT1 siRNA and scramble sequence was used at the concentration of 50nM to intervention C6 cell, using CCK8 method to detect the cells proliferation and viability. The cells invasiveness was detected by tran-swell system. Real-time RT-PCR was used to detect the expression of lnc RNA-MALAT1, miRNA124 and CREB mRNA. Western Blot experiments was used to detect the expression of CREB protein. Results Compared with the control group, after siRNA intervention, the proliferation of C6 cells was decreased（P〈0.05）, and the invasiveness was reduced（P〈0.05）, the expression of MALAT1 gene in C6 cells was decreased, the expression of miRNA124 increased and the expression of CREB mRNA decreased in C6 cells, the expression of CREB protein in each group was in accordance with the expression of mRNA. Conclusion lnc-MALAT1 plays a key role in glioma cells, MALAT1 can increase the transcription and protein expres-sion of CREB gene by regulating the expression of miRNA124, thereby promoting the proliferation of C6 glioma cells.