下调miR-210的表达对U87细胞增殖、侵袭及凋亡的影响

Effects of down-regulation miR-210 expression on proliferation,invasion and apoptosis of U87 cells

目的:探讨miR-210在胶质瘤组织中的表达及下调miR-210对U87细胞增殖、侵袭及凋亡的影响。方法:选取64例择期行手术治疗的胶质瘤患者的胶质瘤组织和30例因脑外伤行内减压术患者的正常脑组织。培养人脑胶质瘤U87细胞,根据转染物不同将细胞分为反义miR-210组、阴性对照组和空白对照组,利用实时荧光定量PCR技术检测不同脑组织及细胞中miR-210及HIF-1α基因表达,MTT比色法检测细胞增殖情况,Transwell法检测细胞迁移及侵袭能力,利用流式细胞术检测细胞凋亡情况,Western blot法检测细胞Bcl-2、Bax、Caspase-3、Caspase-8和Caspase-9蛋白表达。结果:胶质瘤组织中miR-210和HIF-1αmRNA的表达均高于正常脑组织(P<0.05);胶质瘤组织中miR-210和HIF-1αmRNA相对表达量均与肿瘤直径和病理分级有关(P<0.05);反义miR-210组细胞中miR-210和HIF-1αmRNA的表达均低于阴性对照组和空白对照组(P<0.05)。与阴性对照组和空白对照组相比,反义miR-210组细胞在48、72和96 h时光密度值降低,迁移细胞数和侵袭细胞数减少,细胞凋亡率升高(P<0.05)。反义miR-210组细胞中Bcl-2蛋白相对表达量低于阴性对照组和空白对照组,而Bax、Caspase-3、Caspase-8和Caspase-9蛋白相对表达量则升高(P<0.05)。结论:miR-210在胶质瘤组织中呈高表达,下调miR-210表达可抑制细胞增殖、迁移及侵袭能力,促进细胞凋亡发生。

Aim： To investigate the expression of miR-210 in glioma tissue and effects of antisense miR-210 on proliferation,invasion and apoptosis of U87 cells. Methods： The glioma tissue from 64 cases of patients with glioma and normal brain tissue from 30 patients undergoing decompression surgery were collected. The human glioma cells U87 were cultured and divided into antisense miR-210 group,negative control group and blank control group,according to different transfectant. The expressions of miR-210 and HIF-1α genes in different tissues and different transfected groups were detected by real-time PCR. The cell proliferations,migration and invasion as well as cell apoptosis in different transfected groups were detected by MTT,Transwell assay,and flow cytometry,respectively. The expressions of Bcl-2,Bax,Caspase-3,Caspase-8and Caspase-9 proteins in the transfected groups were detected by Western blot. Results： The relative expression levels of miR-210 and HIF-1α mRNA in glioma tissue were higher than the normal brain tissue（ P〈0. 05）. The relative expression levels of miR-210 and HIF-1α mRNA in glioma tissue were related with tumor size and pathological grade（ P〈0. 05）. Compared with the negative control group and blank control group,the relative expression levels of miR-210 and HIF-1α mRNA in antisense miR-210 group and the density values at 48,72 and 96 h were lower,the migration cells and invasion cells reduced,while the cell apoptosis rate increased（ P〈0. 05）. The relative expression level of Bcl-2 protein in antisense miR-210 group was lower than the negative control group and blank control group,while the relative expression levels of Bax,Caspase-3,Caspase-8 and Caspase-9 proteins were higher（ P〈0. 05）. Conclusion： miR-210 is highly expressed in glioma tissue. Down-regulating the expression of miR-210 could inhibit cell proliferation,migration and invasion,and promote apoptosis of U87 cells.