miR-152对HLA-G的作用靶点预测与验证  

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

作　　者：朱晓明[1,2] 韩涛[3] 苏小花[1] 黄琴莉[1] 常春子[1] 姚元庆[2] 

机构地区：[1]第四军医大学唐都医院妇产科,陕西西安710038 [2]解放军总医院妇产科,北京100853 [3]第四军医大学唐都医院骨科,陕西西安710038

出　　处：《现代肿瘤医学》2013年第10期2180-2182,共3页Journal of Modern Oncology

基　　金：国家自然科学基金资助项目(编号:31000660)

摘　　要：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 3'UTR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premiR-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。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.

关 键 词：miR-152 JEG-3细胞 人类白细胞抗原-G 

分 类 号：R730.23[医药卫生—肿瘤] R737.33[医药卫生—临床医学]