运用激光捕获显微切割对肝细胞癌与肝内胆管癌蛋白质组的分析  

Proteomic analysis of hepatocellular carcinoma and cholangiocarcinoma by laser capture microdissection

作　　者：周福平[1] 胡和平[1] 艾建华[1] 陆豪杰[2] 王红阳[1] 吴孟超[1] 

机构地区：[1]第二军医大学东方肝胆外科医院国际合作生物信号转导研究中心,上海200438 [2]复旦大学生物化学研究所,上海200433

出　　处：《第二军医大学学报》2005年第10期1111-1114,共4页Academic Journal of Second Military Medical University

基　　金：国家重点基础研究("973"计划)课题(001CB510205);国家科技攻关计划(2002BA711A023)

摘　　要：目的:分析肝细胞癌和肝内胆管癌蛋白质的表达差异,获取鉴别两者的分子标志物。方法:通过激光捕获显微切割技术分离肝细胞癌和肝内胆管癌的纯细胞,通过双向电泳和质谱的方法检测两者表达的差异蛋白,再通过免疫组化染色的方法证实差异蛋白表达的情况。结果:肝细胞癌和肝内胆管癌图谱分别检测到(1124±110)、(938±90)个蛋白点。分析发现78个差异蛋白点,通过质谱鉴定出28个有意义的蛋白,20个蛋白仅在肝细胞癌表达或表达明显增高,8个蛋白仅在肝内胆管癌中表达或者高表达。同时免疫组化证实BK125H2.1在肝细胞癌中表达强阳性,而肝内胆管癌中缺失表达。结论:激光捕获显微切割是蛋白质组研究中的一个突破性的技术,可以有效地解决组织异质性的问题;本实验检测到的差异蛋白例如BK125H2.1可能成为鉴别肝细胞癌与肝内胆管癌特异性的分子标记物。Objective..To search for the biomarkers for differential diagnosis between hepatocellular carcinoma （HCC） and intrahepatic cholangiocarcinoma （ICC）. Methods： Laser capture microdissection was used to separate hepatocellular carcinoma and intrahepatic cholangiocarcinoma tissues. Proteome alterations were observed using 2-dimensional polyacrylamide gel electrophoresis and electrospray ionisation tandem mass spectrometry. Immunohistochemistry was used to detect the different protein expressions in HCC and ICC tissues. Results： There were 1 124±110 and 938±90 protein spots in the map of HCC and ICC,respectively. Seventy-eight protein spots were discovered and 28 were identified as significant, with 20 expressed or upregulated in hepatocellular carcinoma and 8 expressed or upregulated in intrahepatic cholangiocarcinoma. Immunohistochemistry found that BK125H2.1 expressed in HCC tissue but not in ICC. Conclusion： Laser capture microdissection is a key technique in proteomic study which can effectively resolve the problem of irrelevant tissues. The proteins discovered in this experiment may serve as a biomarker for differential diagnosis of hepatocellular carcinoma and intrahepatic cholangiocarcinoma.

关 键 词：癌 肝细胞 胆管肿瘤 蛋白质组 激光捕获显微切割 

分 类 号：R735.7[医药卫生—肿瘤]