小干扰RNA敲低Dicer对人脑胶质瘤细胞生物学特征的影响

摘要

Objective To study the effect of silencing Dicer by small interference RNA(siRNA) to suppress the global microRNA(miRNAs) expression on the biological characteristics of TJ905 glioblastoma cells. Methods The silencing effect of RNA interference on Dicer expression was evaluated by revers transcription-polymerase chain reaction(RT-PCR), Western blot analysis and immunofluorescence staining.The cell proliferation rate and cell cycle kinetics were detected by MTT assay and flowcytometry respectively, and the cell invasive ability was evaluated by transwell assay. Results The siRNA targeting Dicer suppressed the expression of Dicer in TJ905 cells. Meanwhile, the proliferation activity and invasive ability were significantly enhanced in cells transfected with Dicer siRNA compared to those cells transfected with scrambled siRNA and the control cells. Conclusion Suppression of Dicer expression renders the glioma cells harboring more aggressive phenotype. This preliminary finding suggests that global lower expression of miRNAs may play an oncogenic role.%目的 研究敲低Dicer酶的表达全面抑制徽小RNA(microRNA,miRNA)成熟对TJ905人脑胶质瘤细胞的生物学特征的影响.方法 采用RNA干扰(RNA interference,RNAi)技术敲低Dicer酶的表达,逆转录PCR、Western印迹及免疫荧光检测TJ905细胞转染小干扰RNA(small interferen,siRNA)后Dicer酶表达情况;应用流式细胞术检测、噻唑兰比色分析法试验及Tranwell试验评价细胞生长、增殖和侵袭等生物学特征变化.结果 转染siRNA靶向Dicer酶后,Dicer酶被敲低,S期细胞增多,细胞增殖加速,细胞侵袭生长能力增强.结论 敲低Dicer酶全面抑制miRNAs成熟后,肿瘤细胞表型具有更为恶性变的倾向,由此初步推测全面抑制细胞miRNAs表达有可能促进肿瘤生成.