吲哚美辛对结肠癌细胞成瘤裸鼠作用的蛋白质组学研究  

Proteomic analysis of effects of indomethacin on colorectal cancer xenografts in nude mice

作　　者：张桂英[1] 王郁杰[1] 肖志强[2] 王红梅[1] 程艳丽[1] 

机构地区：[1]中南大学湘雅医院消化科,长沙410008 [2]卫生部肿瘤蛋白质组学重点实验室

出　　处：《中华消化杂志》2006年第3期183-186,共4页Chinese Journal of Digestion

基　　金：国家自然科学基金资助项目(30271516)

摘　　要：目的研究吲哚美辛对人结肠癌细胞系HCT116成瘤裸鼠蛋白质表达谱的影响,寻找吲哚美辛抗结肠癌作用的相关蛋白。方法应用固相pH梯度双向凝胶电泳技术分离吲哚美辛处理组和未处理组HCT116成瘤裸鼠瘤组织的总蛋白,胶体考马斯亮蓝染色,图像扫描获取凝胶电泳图谱后,分析比较两组间差异表达的蛋白质,基质辅助激光解析电离飞行时间质谱测得经胶内胰酶酶解的差异蛋白点的肽质指纹图谱,用Mascot软件查询数据库。结果所获双向凝胶电泳图谱分辨率高、重复性好, 蛋白质点数约1100,吲哚美辛处理组与未处理组的匹配率分别为96．0％和93．6％。两组间共有31个差异表达蛋白点,吲哚美辛处理组有6个蛋白质点表达上调,25个表达下调。获得18张肽质指纹图,经查询数据库初步鉴定12个蛋白质,多数蛋白与肿瘤增殖、浸润、凋亡和肿瘤免疫等相关,包括半乳糖凝集素-1、膜联蛋白及转录因子等。结论吲哚美辛对HCT116结肠癌细胞成瘤裸鼠的作用可能与多种蛋白直接或间接相关,这些蛋白的研究有助于进一步阐明吲哚美辛抗结肠癌作用机制及肿瘤治疗药物新型靶标的发现。Objective To investigate the effect of indomethacin on the protein expression profile of colorectal cancer cell line HCT116 xenografts in nude mice, and to screen the indomethacin anticancer-related proteins. Methods The nude mice with colorectal cancer cell line HCT116 xenograft were treated with or without indomethacin, and then the total proteins extracted from xenografts were separated and visualized by immobilized pH gradient two-dimensional gel （2-DE） electrophoresis. The differential expression proteins between the experimental and control groups were analyzed using PDQuest image analysis software, and identified by peptide mass fingerprinting based on matrix-assisted laser desorption ionization time of flight mass spectrometry and database searching. Results The 2-DE patterns with high resolution and reproducibility were obtained, displayed about 1100 protein spots in each gel, and the matching rate in the two groups was 96. 0% and 93. 6%, respectively. Thirty-one differentially expressed protein spots were found between the two groups, of which 25 spots decreased and 6 increased in the expression levels in the experimental group than in the control group. Eighteen peptide mass fingerprinting maps were obtained, and twelve proteins including galectin-1, armexin and trancription factor, were primarily identified after database searching, most of which were involved in tumor cell proliferation, invasion, apoptosis and immunity. Conclusions Indomethacin exerts its inhibiting effects on colorectal cancer through regulating multiple proteins expression directly or indirectly. The data will be helpful to elucidate the mechanism inhibiting cancer by indomethaein, and to find new targets of chemotherapeutic drugs.

关 键 词：吲哚美辛 结肠癌 蛋白质组学 质谱 

分 类 号：R735.35[医药卫生—肿瘤]