CPEB4对脑胶质瘤细胞增殖、迁移的影响

Effect of CPEB4 on proliferation and migration of human glioma cells

目的:探讨胞质多聚腺苷酸化元件结合蛋白4(CPEB4)对脑胶质瘤细胞增殖、迁移的影响。方法:采用实时荧光定量PCR法检测40例脑胶质瘤组织及对应的癌旁正常组织中CPEB4 mRNA的表达水平。分别将CPEB4 siRNA及无义siRNA转染至脑胶质瘤细胞U87中,Western blot法检测细胞中CPEB4、MMP-9、MMP-2、STAT3、p-STAT3、JAK2和p-JAK2蛋白的表达水平,MTT法检测细胞存活率,细胞划痕实验检测细胞迁移距离,以未处理细胞作对照。使用50μmol/L特异性JAK2抑制剂AG490作用于转染CPEB4 siRNA的U87细胞48 h后,检测细胞存活率及迁移距离,以未处理细胞和AG490处理的U87细胞作对照。结果:脑胶质瘤组织中CPEB4 mRNA表达水平高于癌旁正常组织(P<0.05)。与对照组及无义siRNA组相比,CPEB4 siRNA组细胞CPEB4蛋白表达水平下降(P<0.05),细胞存活率、迁移距离降低(P<0.05),MMP-9、MMP-2、STAT3、p-STAT3、JAK2和p-JAK2蛋白表达水平下降(P<0.05)。与对照组相比,AG490组与AG490+CPEB4 siRNA组细胞存活率和迁移距离均降低(P<0.05);与AG490组相比,AG490+CPEB4 siRNA组细胞存活率与迁移距离降低(P<0.05)。结论:CPEB4在脑胶质瘤组织中高表达,可能通过上调JAK/STAT3信号通路促进脑胶质瘤细胞的增殖与迁移。

Aim：To investigate the effect of CPEB4 on the proliferation and migration of human glioma cells. Methods：The level of CPEB4 mRNA in 40 cases of glioma cancer and paracancerous normal tissue was detected by qRT-PCR.CPEB4 siRNA and nonsense siRNA were transfected into U87 cells for 48 h,the expressions of CPEB4,MMP-9,MMP-2,STAT3,p-STAT3,JAK2 and p-JAK2 were detected by Western blot,cell proliferation was measured by MTT assay,cell scratch test was used to measure the cell migration distance,the proliferation and migration distance of U87 cells transfected with CPEB4 siRNA after 50 μmol/L AG490 treatment for 48 h were detected. The cells without any treatment were the control. Results：The expression of CPEB4 mRNA in glioma tissue was significantly higher than that in paracancerous normal tissue（ P〈0. 05）. Compared with the two control groups,the expression of CPEB4 protein in CPEB4 siRNA group decreased（ P〈0. 05）,cell survival and migration distance were decreased（ P〈0. 05）,and the expressions of MMP-9,MMP-2,STAT3,p-STAT3,JAK2 and p-JAK2 decreased（ P〈0. 05）. Compared with the control group,the proliferation and migration distance in AG490 group and AG490 ＋ CPEB4 siRNA group were significantly reduced（ P〈0. 05）; compared with AG490 group,those in AG490 ＋ CPEB4 siRNA group were significantly reduced（ P〈0. 05）. Conclusion：CPEB4 is highly expressed in glioma tissue,and might promote the proliferation and migration of glioma cells by up-regulating JAK/STAT3 signal pathway.