【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。更多还原

【Abstract】 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 colorectal cancer. Methods The proteins interacting with GPAA1 was screened by co-immunoprecipitation and identified by MALDI-TOF / TOF-MS. Based on bioinformatics 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 colorectal 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), microtubule-associated protein 6( MAP6),beta-1 and 4-galactosyltransferase 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 colorectal cancer cells.更多还原