Zeng Xi, Liao Ai-Jun, Tang Hai-Lin, Yi Lan, Xie Na, Su Qi
Cancer Research Institute, University of South China, Hengyang, Hunan, 421001, P. R. China.
Ai Zheng. 2007 Oct;26(10):1080-4.
BACKGROUND & OBJECTIVE: It is urgent to find the specific or relatively specific biomarkers in the study of gastric carcinoma. This study was to screen human gastric carcinoma-associated antigens by serologic proteome analysis (SERPA).
Total protein were isolated from 10 specimens of human gastric carcinoma by 2-dimensional electrophoresis (2-DE). The immunoblotting imaging films were obtained and the differential reacting protein spots were recognized. Then the differential reacting proteins were obtained in the replica gel by matching analysis, and identified by peptide mass fingerprint (PMF) based on matrix-assisted laser desorption/ionization time of flight mass spectrometry (MALDI-TOF-MS) and database searching.
Fourteen differentially expressed proteins (react only to the patient's serum) were found. Of the 14 proteins, 13 were identified as human gastric carcinoma-associated antigens, including heterogeneous nuclear ribonucleoprotein K (hnRNP K), hnRNP H1, hnRNP H2, cytokeratin 18 (CK18), crystal structure of Fkbp52 C-terminal domain complex with the C-terminal peptide meevd of hot-shock protein 90 (Hsp90), serum vitamin D-binding protein precursor (DBP), selenium-binding protein 1 (SBP1), eukaryotic translation elongation factor 1 delta (eEF-1), ACTB protein, brain creatine kinase (CKB), proteasome 26S ATPase subunit 3 (PSMC3), dimethylarginine dimethylaminohydrolase 1(DDAH-1), and NADH dehydrogenase Fe-S protein 1.
Thirteen human gastric carcinoma-associated antigens were found by SERPA. The results provide foundation for screening molecular biomarkers for the diagnosis, therapy, prognostic evaluation, and prevention of human gastric carcinoma.
在胃癌研究中,迫切需要找到特异性或相对特异性的生物标志物。本研究旨在通过血清蛋白质组分析(SERPA)筛选人胃癌相关抗原。
采用二维电泳(2-DE)从10例人胃癌标本中分离总蛋白。获取免疫印迹成像胶片并识别差异反应蛋白斑点。然后通过匹配分析在复制凝胶中获得差异反应蛋白,并基于基质辅助激光解吸/电离飞行时间质谱(MALDI-TOF-MS)和数据库搜索,通过肽质量指纹(PMF)进行鉴定。
发现14种差异表达蛋白(仅与患者血清反应)。在这14种蛋白中,13种被鉴定为人胃癌相关抗原,包括不均一核核糖核蛋白K(hnRNP K)、hnRNP H1、hnRNP H2、细胞角蛋白18(CK18)、Fkbp52 C末端结构域与热休克蛋白90(Hsp90)的C末端肽meevd的晶体结构、血清维生素D结合蛋白前体(DBP)、硒结合蛋白1(SBP1)、真核翻译延伸因子1δ(eEF-1)、ACTB蛋白、脑肌酸激酶(CKB)、蛋白酶体26S ATP酶亚基3(PSMC3)、二甲基精氨酸二甲胺水解酶1(DDAH-1)和NADH脱氢酶铁硫蛋白1。
通过SERPA发现了13种人胃癌相关抗原。这些结果为筛选用于人胃癌诊断、治疗、预后评估和预防的分子生物标志物提供了基础。