应用蛋白质组学方法筛选结肠癌相关蛋白质  

Screening of colon carcinoma associated proteins by proteomics technique

作　　者：裴海平[1] 朱红[2] 曾亮[3] 李宜雄[1] 

机构地区：[1]中南大学湘雅医院普外科,湖南长沙410008 [2]中南大学湘雅医院肿瘤科,湖南长沙410008 [3]湖南省肿瘤医院病理科,湖南长沙410006

出　　处：《实用肿瘤杂志》2007年第2期112-118,共7页Journal of Practical Oncology

基　　金：湖南省科委课题(04SK3043-1;05SK3007);湖南省卫生厅课题(C2005-009)资助

摘　　要：目的通过比较结肠癌组织与癌旁正常肠组织的差异表达蛋白谱,以期发现结肠癌癌变相关蛋白。方法将结肠癌组织与癌旁正常结肠组织蛋白进行双向凝胶电泳,选择在癌组织中高表达的50个点进行MALDI-TOF质谱分析和生物信息学分析。Western Blot和免疫组化方法验证蛋白的差异表达。结果建立了结肠癌及正常肠组织的双向凝胶电泳图谱,其中癌旁正常结肠组织和结肠癌组织电泳图谱中平均蛋白质点数分别为860±45和980±28;癌旁正常结肠组织和结肠癌组在IEF方向上的平均偏差分别为(0.487±0.11)mm和(0.654±0.11)mm,在SDS-PAGE方向上的平均偏差分别为(0.944±0.12)mm和(1.20±0.22)mm,结肠癌与癌旁正常结肠组织的差异表达蛋白质点数为72.00±12.34。选择在癌组织中高表达的50个点进行质谱分析和生物信息学查询,鉴定了其中有意义的25个点,包括谷胱苷肽S转移酶、肝脂肪酸结合蛋白、热休克蛋白27等。免疫组化方法检测结肠癌组织芯片中GST和HSP27的表达,结果显示,GST在结肠癌中表达的阳性率为73.7%,在正常肠黏膜中表达的阳性率为28.6%,差异有显著性(P<0.01);HSP27在结肠癌中表达的阳性率为73.7%,在正常肠黏膜中表达的阳性率为21.4%,差异有显著性(P<0.01)。结论蛋白质组学方法较好地显示结肠癌组织和癌旁正常组织的差异表达蛋白;其中在结肠癌组织中高表达蛋白HSP27和GST可能作为结肠癌诊断标志物。Objective To find proteins involved in colon carcinogenesis by comparing differentially expressed proteins between colon carcinoma tissues and paired normal tumor-adjacent colon tissues. Methods Total tissue proteins were extracted; differential proteome profiles were established by means of immobilized pH gradient-based two-dimesional polyacrylamide gel electrophoresis （2D-PAGE） and analysed by matrix-assisted laser desorption/ionization time of flight mass spectrometry （ MALDI-TOF-MS ） and bioinformatics. Differentially expressed proteins were observed by immunohistochemical method combined with tissue chip. Results Well-resolved,reproducible 2-DE profiles of human colon carcinoma tissues and paired normal tumor-adjacent colon tissues were obtained. For colon carcinoma tissues,a total of （980~ 28） spots were detected;for paired normal tumor-adjacent colon tissues, （860-1-45） spots were detected. For normal tumor- adjacent colon tissues,the average deviation of spot position was （10. 487± 0.11）mm in IEF direction and （0. 944± 0. 12）mm in SDS-PGE direction; for colon carcinoma tissues the average deviation of spot position was （0. 654 ± 0. 11）mm in IEF direction and （1.20± 0. 22）mm in SDS-PGE direction. Number of differentially expressed proteins was 72.00± 12.34 between colon carcinoma and paired normal tumor-adjacent colon tissues. Fifty differentailly expressed proteins were analysed by mass spectrometry and bioinformatics, 25 of them were well characterized including glutathione S-transferase （GST）,hepatic fatty acid-binding protein, heat shock protein 27 （HSP27）, ect. Positive rate of GST exprssion in carcinoma group （73. 7%） was significantly higher than that in normal group （28.6%） （P〈（0. 01）; positive rate of HSP27 exprssion in carcinoma group （73.7%） was significantly higher than that in normal group（21.4%） （P〈（0. 01）. Conclusion Proteomic technique is a good method to find differentially expressed proteins between colon carcino

关 键 词：结肠肿瘤/病理学 热休克蛋白质类/分析 蛋白质组学 谷胱苷肽转移酶 电泳 凝胶 双向 载体蛋白质类/分析 

分 类 号：R735.3[医药卫生—肿瘤]