摘要
目的应用差异蛋白质组学方法筛查血清中与肝癌转移相关的蛋白质分子。方法收集具有相同疾病背景原发性肝癌(肝内转移和未转移)患者以及正常人的血清(每组各20例),经过去高丰度蛋白预处理后,进行双向电泳(2-DE)分析,筛选出肝内转移肝癌与未转移肝癌相比较有显著性差异的蛋白质斑点,进行MALDI-TOF/TOF质谱鉴定;使用Western blotting验证蛋白质组分析的结果。结果对3组血清的双向电泳图谱分析,与正常人和未转移肝癌患者的血清相比,在肝内转移肝癌血清中共筛选出12个差异大于2倍的蛋白质斑点,MALDI-TOF/TOF成功鉴定出3个表达下调的蛋白质[包括Inter-alpha(globulin)inhibitor H4、Complement factor H-related protein1precursor(FHR-1)、apolipoprotein E]和一个上调蛋白(Cytokeratin9,CK9)。Western blotting对所有样本血清中的CK9进行了检测,结果与蛋白质组分析结果相一致。结论肝癌的转移与多种蛋白表达改变相关,其中CK9的高表达可能与肝癌的转移有关,可能作为肝癌转移相关的潜在血清标志物。
Objective To screen the serum proteins associated with the metastasis of hepatocellular carcinoma （HCC） using a comparative proteomic approach. Methods The serum samples of HCC patients with the same disease background were divided into metastatic （n=20） and non-metastatic （n=20） groups. The proteins extracted from the patients and 20 normal subjects, after depletion of the highly abundant proteins, underwent two-dimensional gel electrophoresis （2-DE）. Comparative analyses of the 2-DE protein patterns between the 3 groups were conducted using a computerized image analysis system. The proteins with statistically significant differential expression between the metastatic and non-metastatic patients were identified by mass spectrometry. Western blotting was performed to examine the differential expression of the candidate proteins. Results Four protein spots were identified by mass spectrometry among the 12 differentially expressed protein spots in the serum samples of HCC patients with intrahepatic metastasis, and confirmed by searching in MASCOT database. Of the 4 proteins, cytokeratin 9 （CK9） was up-regulated by 2 folds, and inter-alpha （globulin） inhibitor H4, complement factor H-related protein 1 precursor （FHR-1）, and apolipoprotein E were down-regulated by 2 folds. CK9 was found to be specifically over-expressed in the metastatic group in comparison with the non-metastatic group, as confirmed by Western blotting. Conclusion The metastasis of HCC might be correlated to the specific variation of protein expression profiles. The overexpression of CK9 may play a crucial role in HCC metastasis, and can be used as a potential serum marker for predicting HCC metastasis.
出处
《南方医科大学学报》
CAS
CSCD
北大核心
2009年第9期1775-1778,共4页
Journal of Southern Medical University
基金
国家重点基础研究发展计划(973计划)(2009CB522404)
国家十一五科技重大专项课题(2008ZX10209)
2007-2009年度卫生部部属(管)医院临床学科重点项目
广东省科技计划项目重大专项(2007A032000001)
中国博士后科学基金(20070420794)
关键词
肝细胞癌
转移
蛋白质组学
血清
hepatocellular carcinoma
metastasis
proteomics
serum
