miR-451对胶质瘤细胞株A172生物学特征的影响

Influence of miR -451 on biological characters of glioma cell line A172

目的 研究miR-451对胶质瘤细胞株A172生物学特征的影响.方法 合成寡核苷酸miR-451拟似物（miR-451 mimics）转染胶质瘤细胞以上调miR-451的表达,real-time PCR检测转染后miR-451的表达,Western印迹法检测目标蛋白表达,应用流式细胞术、MTT法评价细胞生长和增殖的生物学特征变化.结果 转染miR-451 mimics后,real-time PCR检测提示miR-451表达升高,Western blot结果显示癌基因C-myc表达下降＞63.6%,细胞周期正向调节因子CDK2、CDK4、Cyclin D1和Cyclin E表达下降;细胞周期分析表明miR-451 mimics组进入G0/G1期的细胞数较对照组增多达17.4%,出现G0/G1期阻滞;MTT法分析显示miR-451 mimics组细胞生长受抑.结论 miR-451可以抑制人脑胶质瘤细胞生长和增殖能力,具有抑瘤作用.

Objective To investigate the influence of miR -451 on biological characters of gliomas cells A172. Method The oligonucleotide of miR -451 mimics was transfected into human glioblastoma cell line A172. The expression of miRNA -451 was identified by real - time PCR after transfection, and targeting protein expression was evaluated by Western blot. The cell proliferation was determined by flow cytometry and MTT assay. Moreover,cell cycle molecules were detected by Western blot analysis. Results The miR -451was markedly up -regulated after transfection of miRNA -451 mimics group. As compared with control and scramble group, the cell cycle was significantly arrested over 17.4% in G0/G1 phase by flow cytometry assay,meanwhile the protein expression of C - myc and regulatory molecules for cell cycle progression including CDK2, CDK4, Cyclin D1 and Cyclin E were all down -regulated. The proliferation was significantly decreased by the MTT assay at the same time. Conclusions MiR - 451 could inhibit the proliferation activity of A172 glioma cell. It suggests that miR -451 is a tumor suppressor gene in glioma.