Li Cui, Chen Zhu-Chu, Xiao Zhi-Qiang, Wu Xiao-Ying, Zhan Xian-Quan, Li Mao-Yu, Feng Xue-Ping, Zhang Xiao-Peng, Li Jian-Ling, Chen Ping, Liang Song-Ping
Medical Research Center, Xiangya Hospital,Central South University, Changsha, Hunan, 410008, P.R.China.
Ai Zheng. 2004 Jan;23(1):28-35.
BACKGROUND & OBJECTIVE: Carcinogenesis of lung squamous carcinoma is a complex process involving multiple events and steps. Though some molecular pathogenesis studies on human lung cancer have been undertaken successfully in gene (DNA) and transcription (mRNA) levels, the carcinogenic mechanism is still unclear. At present, there is no special molecular marker for early-stage diagnosis and prognosis evaluation. The objective of this study was to establish 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 proteins.
The total proteins of human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue were separated by immobilized pH gradient (IPG)-based two-dimensional gel electrophoresis (2-DE). The differential expression proteins were analyzed using image analysis software, then identified using mass spectrometry and database searching.
(1) For tumor tissues, the average protein spots of 3 gels were 1349+/-67; and 1235+/-48 spots were matched with the average matching rate of 91.5%. For control, the average protein spots of 3 gels were 1297+/-73; and 1183+/-56 spots were matched with the average matching rate of 91.2%. The average position deviation of matched spots was 0.873+/-0.125 mm in IEF direction, and 1.025+/-0.213 mm in SDS-PAGE direction. (2) A total of 1069+/-45 spots were matched between the electrophoretic maps of 15 human lung squamous carcinoma tissue and paired normal tumor-adjacent bronchial epithelial tissue. Forty differential proteins were identified by peptide mass fingerprint(PMF), some proteins were the products of oncogenes, and the others involve in the regulation of cell cycle and signal transduction.
In this study, the well-resolved, reproducible 2-DE patterns of human lung squamous carcinoma and adjacent normal bronchial epithelial tissues were established and certain differential proteins were characterized. These data will be helpful for screening the biomarker to further study on human lung squamous carcinoma.
肺鳞癌的发生是一个涉及多个事件和步骤的复杂过程。尽管在基因（DNA）和转录（mRNA）水平上已经成功开展了一些关于人类肺癌的分子发病机制研究，但致癌机制仍不清楚。目前，尚无用于早期诊断和预后评估的特异性分子标志物。本研究的目的是建立人肺鳞癌组织及配对的正常肿瘤旁支气管上皮组织的高分辨率、可重复性二维电泳图谱，并鉴定差异表达蛋白。
采用基于固定化pH梯度（IPG）的二维凝胶电泳（2-DE）分离人肺鳞癌组织及配对的正常肿瘤旁支气管上皮组织的总蛋白。使用图像分析软件分析差异表达蛋白，然后通过质谱和数据库检索进行鉴定。
（1）肿瘤组织方面，3块凝胶的平均蛋白点数为1349±67；匹配的平均点数为1235±48，平均匹配率为91.5%。对照组织方面，3块凝胶的平均蛋白点数为1297±73；匹配的平均点数为1183±56，平均匹配率为91.2%。匹配点在等电聚焦（IEF）方向的平均位置偏差为0.873±0.125mm，在十二烷基硫酸钠-聚丙烯酰胺凝胶电泳（SDS-PAGE）方向为1.025±0.213mm。（2）15例人肺鳞癌组织及配对的正常肿瘤旁支气管上皮组织的电泳图谱之间共匹配1069±45个点。通过肽质量指纹图谱（PMF）鉴定出40种差异蛋白，其中一些蛋白是癌基因的产物，其他蛋白参与细胞周期调控和信号转导。
本研究建立了人肺鳞癌及相邻正常支气管上皮组织分辨率良好、可重复的2-DE图谱，并鉴定了某些差异蛋白。这些数据将有助于筛选生物标志物，以进一步研究人肺鳞癌。
