目的采用以肿瘤免疫学与蛋白质组学研究技术有机地结合为基础的血清蛋白质组学研究体系(SERPA)筛选肺腺癌相关抗原。方法应用SERPA对4例人肺腺癌组织进行研究,分析肺腺癌组织总蛋白分别与肺腺癌患者的自身血清以及正常对照血清反应的W estern b lot图谱,识别鉴定差异反应的蛋白质。结果获得了分辨率较高的人肺腺癌组织与患者的自身血清以及正常对照血清的W estern b lot图谱,共识别27±5个差异反应的蛋白质。对其中的部分差异蛋白质点进行了肽质指纹图分析,鉴定出一些与细胞生长增殖、细胞代谢、细胞周期调控、信号转导等有关的肺腺癌相关抗原。结论本研究为进一步筛选用于肺腺癌诊断、治疗和预后评估的肺腺癌分子标志物奠定了坚实的基础。
Objective： To establish the two-dimensional electrophoresis profiles with high resolution and reproducibility from human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue, and to identify differential expression tumor-associated proteins by using proteome analysis. Methods： Comparative proteome analysis with 20 human lung squamous carcinoma tissues and the paired normal bronchial epithelial tissues adjacent to tumors was carried out. The total proteins of human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue were separated by means of immobilized pH gradient-based two-dimensional gel electrophoresis （2-DE） and silver staining. The differential expression proteins were analyzed and then identified by matrix-assisted laser desorption/ionization time of flight mass spectrometry （MALDI-TOF-MS）. Results： （1） Well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were obtained. For tumor tissue, average spots of 3 gels were 1567±46, and 1436±54 spots were matched with an average matching rate of 91.6%. For control, average spots of 3 gels were 1349±58, and 1228±35 spots were matched with an average matching rate of 91.03%. The average position deviation of matched spots was 0.924±0.128 mm in IEF direction, and 1.022±0.205 mm in SDS-PAGE direction; （2） A total of 1178±56 spots were matched between the eleetrophoretie maps of 20 human lung squamous carcinoma tissues and paired normal tumor-adjacent bronchial epithelial tissues. Seventy-six differentially expressed proteins were screened; （3） Sixty-eight differential proteins were identified by PMF, some proteins were the products of oneogenes, and others involved in the regulation of cell cycle and signal transduetion; （4） In order to validate the reliability of the identified results, the expression of 3 proteins mdm2, c-jun and EGFR, which was correlated with lung squamous carcino
LI CuiTANG Can＇eDUAN ChaojunYI HongXIAO ZhiqiangCHEN Zhuchu
