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.
出处
《第二军医大学学报》
CAS
CSCD
北大核心
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)
关键词
癌
肝细胞
胆管肿瘤
蛋白质组
激光捕获显微切割
carcinoma,hepatocellular
biliary tract neoplasms
proteome
laser capture microdissection