HSPC238对HMOX1、RPS27a、MT2A、UBB调节的初步研究  

A preliminary study on regulation of HMOX1,RPS27a,MT2A and UBB by HSPC238

作　　者：谭家余[1] 黄湘[1] 陈敬林[1] 钟裕恒 

机构地区：[1]南方医科大学第三临床医学院南方医科大学附属中山博爱医院,中山528403

出　　处：《中国免疫学杂志》2016年第4期509-512,共4页Chinese Journal of Immunology

基　　金：国家自然科学基金(81101534);广东省自然科学基金(S2012010010824);广东省医学科研基金(A2013875)项目资助

摘　　要：目的:探讨HSPC238过表达或低表达对目标蛋白(HMOX1、MT2A、UBB、RPS27a)的调节作用及其调节途径。方法:分别构建HSPC238的干扰载体及高表达载体,脂质体法转染HEPG2细胞株,RT-PCR、Western blot检测HSPC238及目标蛋白的mRNA、蛋白的表达量;进一步用目标蛋白的高表达质粒分别转染低表达及高表达HSPC238的HEPG2细胞株,实验组另以蛋白酶体抑制剂MG132处理细胞,镍柱纯化目标蛋白,以HSPC238做免疫印迹,检测目标蛋白累积情况。结果:外源性干扰HSPC238后HMOX1、MT2A、RPS27a表达下调较明显,外源性过表达HSPC238后MT2A、UBB、RPS27a表达水平明显上调;目标蛋白的高表达质粒分别转染低表达及高表达HSPC238的HEPG2细胞株,实验组以蛋白酶体抑制剂MG132处理细胞后,与对照组比较,泛素化的目标蛋白条带明显增加。结论:过表达HSPC238可上调MT2A、UBB、RPS27a的表达,干扰HSPC238可下调HMOX1、MT2A、RPS27a的表达;HSPC238可能通过泛素蛋白酶体途径(Ubiquitin-proteasome pathway,UPP)对目标蛋白发挥调节作用。Objective： To investigate the effect of HSPC238 overexpression or low expression on the regulation of the target protein（ HMOX1,MT2 A,UBB,RPS27a） and the regulation pathways. Methods： To construct the interference vector and overexpression vector of HSPC238 respectively,transfected the HEPG2 cell lines by the liposome method,detect the expression of mRNA and protein of the HSPC238 and the target proteins by the RT-PCR and Western blot,further to transfer the overexpression plasmids of the target proteins into the HEPG2 cell lines which had been transfected with interference vector and overexpression vector of HSPC238,the experimental group cell lines were treated with proteasome inhibitor MG132,to purify the target proteins with nickel column,do immunoblotting with HSPC238 to detect the accumulation situation of the target proteins. Results： The HMOX1,MT2 A,RPS27a were downregulated obviously when the HSPC238 was interfered exogenous; and the MT2 A,UBB,RPS27 a were up-regulated after the HSPC238 overexpressed. The overexpression plasmid of target proteins were transfected into the HEPG2 cell lines which have been transfected with interference vector and overexpression vector of HSPC238,compared with the control group,the target protein band in the experimental group was significantly increased after treatment with the proteasome inhibitor MG132. Conclusion： The HSPC238 overexpression can upregulate the expression of MT2 A,UBB and RPS27 a,and interfering HSPC238 can downregulate the expression of HMOX1,MT2 A and RPS27a; the HSPC238 target protein may play a regulatory role through the UPP pathway; the HSPC238 may play a regulatory role on the target proteins through the UPP pathway.

关 键 词：HSPC238 泛素蛋白酶体途径 目标蛋白 

分 类 号：R392.1[医药卫生—免疫学]