Lee I-Neng, Chen Chien-Hung, Sheu Jin-Chuan, Lee Hsuan-Shu, Huang Guan-Tarn, Chen Ding-Shinn, Yu Chen-Yin, Wen Chu-Ling, Lu Fung-Jou, Chow Lu-Ping
Graduate Institute of Biochemistry and Molecular Biology, College of Medicine, National Taiwan University, Taipei, Taiwan.
Proteomics. 2006 May;6(9):2865-73. doi: 10.1002/pmic.200500488.
Although the significant risk factors for hepatocellular carcinoma (HCC) are well known from epidemiological studies, diagnosis of this disease at an early stage is difficult, and HCC remains one of the leading causes of cancer death worldwide. Thus, to identify any useful HCC-related biomarkers is still a need. We performed SELDI-TOF MS to identify differentially expressed proteins in HCC serum using weak cation exchange protein chips. Protein characterization was performed by 2-DE separation and nano flow LC-MS/MS. A total of 55 sera were collected from HCC patients and compared with those from 48 patients with chronic hepatitis and 9 healthy individuals. A candidate marker of about 8900 Da was detected as differentially expressed in patients with chronic hepatitis C and hepatitis C virus (HCV)-related HCC. We identified this differentially expressed protein as complement C3a. The expression of C3a in HCC sera was further validated by PS20 chip immunoassay and Western blotting. Complement C3a was found to be elevated in patients with chronic hepatitis C and HCV-related HCC. The combination of SELDI-TOF MS and 2-DE provides a solution to identify disease-associated serum biomarkers.
尽管从流行病学研究中已熟知肝细胞癌（HCC）的重要风险因素，但早期诊断这种疾病仍很困难，HCC仍是全球癌症死亡的主要原因之一。因此，仍需要鉴定任何有用的HCC相关生物标志物。我们使用弱阳离子交换蛋白芯片进行表面增强激光解吸电离飞行时间质谱（SELDI-TOF MS），以鉴定HCC血清中差异表达的蛋白质。通过二维电泳（2-DE）分离和纳升流液相色谱-串联质谱（nano flow LC-MS/MS）进行蛋白质表征。共收集了55份HCC患者的血清，并与48例慢性肝炎患者和9名健康个体的血清进行比较。在丙型肝炎和丙型肝炎病毒（HCV）相关的HCC患者中，检测到一种约8900 Da的候选标志物差异表达。我们将这种差异表达的蛋白质鉴定为补体C3a。通过PS20芯片免疫测定和蛋白质印迹进一步验证了C3a在HCC血清中的表达。发现补体C3a在丙型肝炎和HCV相关的HCC患者中升高。SELDI-TOF MS和2-DE的结合为鉴定疾病相关的血清生物标志物提供了解决方案。
