双向电泳分析CCR5对二氢叶酸还原酶表达的抑制作用  

Analysis of inhibitory effect of CCR5 on dihydrofolate reductase expression by two-dimensional electrophoresis

作　　者：邱磊[1] 丁力[1] 张冉[1] 冯皓[1] 郭葆玉[1] 

机构地区：[1]第二军医大学药学院生化药学教研室,上海200433

出　　处：《第二军医大学学报》2008年第7期768-772,共5页Academic Journal of Second Military Medical University

基　　金：国家自然科学基金(30371349)~~

摘　　要：目的:采用双向电泳为基础的蛋白质组学的研究手段,发现趋化因子受体5被配体激活后的功能相关蛋白质。方法:构建稳定表达CCR5的HEK-293细胞株。用RANTES刺激12 h后,用双向电泳方法进行分析。在以PDQuest图像分析软件进行分析后,对表达有差异的蛋白质进行MOLDI-TOF MS/MS质谱鉴定,并对鉴定的蛋白作RT-PCR初步分析。结果:流式细胞仪检测结果表明HEK-293细胞稳定表达CCR5,在受RANTES刺激后作双向电泳分析并经质谱鉴定后,发现二氢叶酸还原酶表达降低,与RT-PCR检测结果相一致。结论:经蛋白质学组方法分析,发现CCR5被其配体RANTES 激活后抑制了二氢叶酸还原酶的表达。Objective： To identify the differentially expressed proteins after activation of CCR5 by its ligand using twodimensional electrophoresis. Methods： The HEK-293 cells stably expressing CCR5 were constructed. The global protein analysis in 293-CCR5 stables and mock cells was performed after treatment with RANTES. After analysis with the PDQuest image software, the differential spots were identified by MOLDI-TOF MS/MS. RT-PCR was performed to further analyze the changes at mRNA level. Results： Flow cytometry revealed that HEK-293 cells stably expressed CCRS. In this study, we analyzed the proteomic results to reveal the proteins which were significantly modulated by the activation of CCR5 with RANTES. Expression of dihydrofolate reductase was found in HEK-293 cells after RANTES treatment, which was confirmed by RT-PCR at the mRNA level. Conclusion.. This proteomic study suggests that CCR5 activated by its ligand RANTES inhibits the expression of dihydroiolate reductase.

关 键 词：双向凝胶电泳 RANTES CCR5受体 四氢叶酸脱氢酶 

分 类 号：R345.4[医药卫生—基础医学]