miR-122a 抑制人喉癌细胞株Hep2 细胞增殖的研究

目的:研究miR-122a 对人喉癌细胞株Hep2 细胞增殖、细胞周期以及相关蛋白表达量的影响。方法:人喉癌细胞株Hep2 细胞分别转染miR-122a 寡聚核苷酸(A 组)和miR-122a inhibitor(阻遏物)(B 组),同时设立阻遏物阴性对照( inhibitor negative control,miR-NC inhibitor)(C 组)和空白对照(D 组)。用RT-PCR、MTT 法、流式细胞仪和Western blot 技术评价各组细胞增殖和细胞周期的生物学特征。结果:转染miR-122a 寡聚核苷酸后,Hep2 细胞中miR-122a 表达显著增加。同D 组相比,A 组细胞增殖水平明显受到抑制,miR-122a 寡聚核苷酸能够有效诱导Hep-2 细胞周期阻滞在G1/ G0 期,A 组细胞分裂周期蛋白42 表达水平明显下调,细胞周期调控因子蛋白CDK4 及细胞周期素D1 蛋白表达水平明显降低。结论:miR-122a 寡聚核苷酸能够显著抑制喉癌细胞Hep2 的增殖,miR-122a 是潜在的人喉癌细胞基因治疗的候选靶点。

Impact of miR-122a on inhibition proliferation of laryngeal carcinoma cell line Hep2

Objective:To study the effect of miR-122a on inhibition proliferation of laryngeal carcinoma cell line Hep-2. Methods: The oligomucleotide of miR-122a was transfected into laryngeal carcinoma cell line Hep2 cells,which were devided into three groups of A(miR-122a transfection),B(miR-122a inhibitor),C(miR-122a-NC inhibitor) and group of D(blank control).The expression of miR-122a was defected by RT-PCR,and relevant protein expression was evaluated by Western blot.The cell proliferation and cell cycle were determined by MTT assy and flow cytometry,respectively. Results: Compared to group D,miR-122a expression in Hep2 cells was obviously elevated atter miR-122a-transfected.The proliferation of Hep2 cells in group A was significantly inhibited and the cell cycle arrested at G1/ G0 phase. The protein expression of CDC42 was downregulated with decreased expressions of CDK4 and cyclin D1 in group A.Conclusion: miR-122a inhibits the proliferation activity of Hep2 cells,suggesting that miR-122a can be taken as a potential candidate for gene therapy of laryngeal carcinoma.