Ying Wantao, Zhang Kaitai, Qian Xiaohong, Xie Ling, Wang Jie, Xiang Xiaoqiong, Cai Yun, Wu Dechang
Department of Genomics and Proteomics, Beijing Institute of Radiation Medicine, Beijing, P.R. China.
Proteomics. 2003 Jan;3(1):64-72. doi: 10.1002/pmic.200390010.
To probe the mechanism of carcinogenesis of lung cancer at the molecular level and to find potential protein markers involved in the early phase of tumorgenesis, differential proteome analysis on primary passage cell line R15H, and early transformed cell line R15H20 derived from (238)Pu alpha-particle irradiation of human papillomavirus (HPV) 18-immortalized human bronchial epithelial cell line (BEP2D), was carried out using two-dimensional electrophoresis (2-DE) and peptide mass fingerprinting (PMF) with matrix-assisted laser desorption/ionisation-time of flight mass spectrometry. Image analysis and Student's t-test (p < 0.05) showed that three protein spots were only expressed in R15H, intensities of 43 protein spots on the gels were altered between R15H and R15H20. Two of the three spots that were only expressed in R15H were identified as high mobility group protein 1. Two proteins decreased in abundance in R15H20 were identified as maspin precursor, a tumor suppressor and aminoacylase-1. Ornithine aminotransferase and peptidyl-prolyl cis-trans isomerase A that were increased in R15H20, were also identified. Relationships between these differentially expressed proteins and the carcinogenesis mechanism of lung cancer are discussed. The protein expression profile of the R15H cell line was also constructed during the study as a reference map for further comparative proteome analysis of the irradiation induced BEP2D cell line. Of the 90 spots analyzed with PMF in the 2-DE gel of R15H cell line, 50 proteins were identified by searching the nonredundant protein database SWISS-PROT/TrEMBL.
为了从分子水平探究肺癌的致癌机制,并寻找肿瘤发生早期阶段涉及的潜在蛋白质标志物,我们利用二维电泳(2-DE)和基质辅助激光解吸电离飞行时间质谱的肽质量指纹图谱(PMF)技术,对人乳头瘤病毒(HPV)18永生化人支气管上皮细胞系(BEP2D)经(238)Puα粒子照射后的原代传代细胞系R15H和早期转化细胞系R15H20进行了差异蛋白质组分析。图像分析和学生t检验(p < 0.05)显示,有三个蛋白质斑点仅在R15H中表达,凝胶上43个蛋白质斑点的强度在R15H和R15H20之间发生了改变。仅在R15H中表达的三个斑点中有两个被鉴定为高迁移率族蛋白1。在R15H20中丰度降低的两种蛋白质被鉴定为肿瘤抑制因子maspin前体和氨基酰化酶-1。在R15H20中表达增加的鸟氨酸氨基转移酶和肽基脯氨酰顺反异构酶A也被鉴定出来。本文讨论了这些差异表达蛋白质与肺癌致癌机制之间的关系。在研究过程中还构建了R15H细胞系的蛋白质表达谱,作为进一步对比分析辐射诱导的BEP2D细胞系蛋白质组的参考图谱。在R15H细胞系的2-DE凝胶中,用PMF分析的90个斑点中,通过搜索非冗余蛋白质数据库SWISS-PROT/TrEMBL鉴定出了50种蛋白质。