过表达和抑制miR-133a对JEG-3中HLA-G表达的影响

Overexpression and specific inhibition of miR-133a regulate the expression of HLA-G in human choriocarcinoma JEG-3 cell lines

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-133 a in JEG-3 cells. Methods The trophblastic JEG-3 cells were transfected with pre-miR-133 a or pre-miR-133 a inhibitor. miR-133 a 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-133 a,miR-133 a significantly increased,and HLA-G mRNA and protein significantly decreased,compared with cells treated with pre-miR-133 a inhibitor（ P〈0. 05）. Conclusion miR-133 a 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.