miR-152对HLA-G的作用靶点预测与验证

目的：预测miR-152分子对HLA-G的作用靶点并进行实验验证.方法：采用PITA软件分析miR-152与HLA-G的相互作用位点,构建含有该作用位点的HLA-G 3UTR真核表达质粒pMIR-UTR.转染实验分为3组：pMIR-UTR与pre-miR-152共转染JEG-3滋养细胞（实验组）,仅转染pMIR-UTR质粒（空白对照）,共转染pMIR-UTR与pre-miR-control分子（阴性对照）,所有细胞均转染pMIR-β-gal质粒以标准化实验背景,转染48h后进行荧光素酶检测.结果：JEG-3细胞中HLA-G的3＇UTR存在与miR-152的种子序列完全互补的结构.荧光素酶报告系统结果显示：与空白对照组和阴性对照组相比,转染pre-miR-152的实验组其荧光素酶活性显著降低（P＜0.05）.结论：miR-152可以通过作用于HLA-G的3 ＇UTR区从而抑制HLA-G的表达.

The prediction and verification of miR-152 target site in HLA-G

Objective：To predict and verify miR - 152 target site in HLA - G. Methods：The prediction of miRNA - 152 target site in HLA - G was performed using PITA software. Lueiferase reporter vector containing miRNA - 152 target site in HLA - G 3i_ITR was constructed. For the test group, transfections of JEG - 3 cells were performed with pMIR - UTR and pre - miR - 152. For the bland control group, transfections were performed with pMIR - UTR but no pre - miR - 152. For the negative control group, transfections were performed with pMIR - UTR and pre - miR - con- trol. All cells were also transfected with pMIR - β -gal for normalizing variability. After 48h, the cells were measured for luciferase and β - gal activity. Results：There was a target sequence of miR - 152 in the 3＇ UTR of HLA - G. The lueiferase reporter assay showed a significant decrease of luciferase activity in the test group compared with the con- trols （P 〈 0.05）. Conclusion： miR- 152 leads to reduced expression of HLA - G through interaction with HLA- G 3＇ UTR.