过表达和抑制miR-133a对JEG-3中HLA-G表达的影响

摘要

目的 研究微小miR-133a在人绒癌细胞株(JEG-3)中对人类白细胞抗原G(HLA-G)的表达调控作用.方法 用脂质体包裹化学合成的miR-133a前体分子并转染JEG-3,从而过表达及抑制miR-133a,逆转录-聚合酶链反应技术(RT-PCR)检测转染后miR-133a及HLA-G在mRNA水平的表达情况.Western blot法检测转染miR-133a后细胞中HLA-G蛋白水平的表达.结果 JEG-3细胞中抑制miR-133a后HLA-G在mRNA及蛋白水平均表达增加;在JEG-3细胞中过表达miR-133a后HLA-G在mRNA及蛋白水平均表达下降,差异有统计学意义(P＜0.05).结论 在JEG-3细胞中miR-133a能够负性调控HLA-G的表达,进一步验证了miR-133a与自然流产、子痫前期等疾病的相关性,并为进一步探索相关疾病机制及临床诊治提供了一定的实验基础.%Objective To investigate the regulation of HLA-G expression by miR-133a in JEG-3 cells.Methods The trophblastic JEG-3 cells were transfected with pre-miR-133a or pre-miR-133a inhibitor.miR-133a mRNA,HLA-G mRNA and HLA-G protein were detected in the transfected JEG-3 cells using RT-PCR and Western blot.Results In cells transfected with pre-miR-133a,miR-133a significantly increased,and HLA-G mRNA and protein significantly decreased,compared with cells treated with pre-miR-133a inhibitor (P ＜ 0.05).Conclusion miR133a down regulates HLA-G expression at the transcription level.Low expression of HLA-G is expected to decrease its immune tolerance effect.This process may be involved in development of immune mediated pathological pregnancy.