免疫共沉淀联合质谱分析筛选结直肠癌组织中GPAA1相互作用蛋白的初步研究  

Screening of proteins interacting with GPAA1 in colorectal cancer by co-immunoprecipitation and MALDI-TOF/TOF-MS

作　　者：陈光[1,2] 李世拥[2] 安萍[2] 蔡慧云[2] 

机构地区：[1]第三军医大学研究生管理大队,重庆400032 [2]北京军区总医院普通外科

出　　处：《中华普外科手术学杂志（电子版）》2014年第1期54-56,共3页Chinese Journal of Operative Procedures of General Surgery(Electronic Edition)

基　　金：国家自然科学基金资助项目(30772118)

摘　　要：目的通过筛选结直肠癌组织中GPAA1相互作用蛋白以探讨GPAA1及其相互作用蛋白在结直肠癌增殖、侵袭、转移过程中的作用。方法利用免疫共沉淀技术筛选新鲜结直肠癌原发灶组织标本中GPAA1相互作用蛋白;利用MALDI-TOF/TOF-MS质谱分析技术分析鉴定GPAA1相互作用蛋白,质谱数据用Mascot软件在NCBInr数据库内进行检索,通过生物信息学分析GPAA1及其相互作用蛋白在结直肠癌增殖、侵袭、转移过程中的作用。结果通过免疫印迹检测发现结、直肠癌原发灶组织中均有GPAA1蛋白表达,大小在70 kD和62 kD之间。质谱分析筛选出5个与GPAA1相互作用蛋白,包括:PHD finger protein 10(PHF10)、促凋亡蛋白Bcl-2修饰因子、Bcl-2-related protein A1(Bcl-2A1)、Microtubule-associated protein 6(MAP6)、β1,4-半乳糖基转移酶。结论利用免疫共沉淀联合质谱分析技术成功筛选出GPAA1相互作用蛋白,GPAA1可能通过Bcl-2家族蛋白相互作用,从而阻断程序性凋亡过程促进结直肠癌细胞增殖,生长。Objective To screen the proteins which interact with GPAA1 in colorectal cancer, and to investigate the impact of interaction of GPAA1 and screened proteins on the proliferation, invasion, and metastasis of coloreetal cancer. Methods The proteins interacting with GPAA1 was screened by eo-immunoprecipitation and identified by MALDI-TOF/TOF-MS. Based on bioinformaties by Mascot software and NCBInr database, we investigated the role of these proteins in the proliferation, invasion, and metastasis of colorectal cancer. Results Results of western blotting showed that GPAA1 overexpressed in primary eolorectal cancer tissues, with the molecular weight from 70 kD to 62 kD. Five novel proteins interacted with GPAA1 were screened by MALDI-TOF/TOF-MS, including PHD finger protein 10 （PHF10）, Bcl-2-modifying factor （ Bmf）, Bcl-2-related protein A1 （ Bcl-2A1 ）, microtubnle-associated protein 6 （MAP6） , beta-1 and 4-galactosyhransferase 4. Conclusions Proteins interacted with GPAA1 could be screened by combined application of co-immunoprecipitation and MALDI-TOF/TOF-MS. GPAA1 might regulate Bcl-2 family proteins expression and induce apoptosis of eolorectal cancer cells.

关 键 词：结直肠肿瘤 糖基磷脂酰肌醇类 免疫沉淀法 

分 类 号：R735.3[医药卫生—肿瘤]