PTPα高表达致NIH3T3细胞恶变早期相关基因的筛选与鉴定  

Screening and Identification of Tumor Related Genes in Early Stages of Transformed NIH3T3 Cells Associated with Overexpressed PTPα

作　　者：杨劲松[1] 郑新民[2] 陈诗书[1] 

机构地区：[1]上海第二医科大学生化教研室,分子生物学实验室,人类基因治疗研究中心,上海200025 [2]美国康奈尔大学,纽约14850

出　　处：《生物化学与生物物理学报》2002年第5期601-607,共7页

摘　　要：利用已建立的蛋白质酪氨酸磷酸酶α(PTPα)诱导表达模型筛选NIH3T3细胞中PTPα诱导表达前后的差别表达基因 ,有利于探索肿瘤早期形成的机制。诱导PTPα表达 2 4h ,用差异显示逆转录PCR获得 6 5条差异片段 ,利用生物信息学方法分析这些差异片段 ,发现其中含有 2 9种已知基因、1 2种已知EST、6种未知EST ;对已知基因进行功能查询和分析 ,发现它们分别与信号转导、细胞骨架及粘附、转录与翻译、能量代谢、凋亡及核糖体蛋白质相关 ,其中G蛋白基因、TCR基因、类Trx基因、小窝蛋白 1 (caveolin 1 )基因经RT PCR及Northern印迹实验证实了差异表达的真实性。结果表明 ,PTPα诱导表达 2 4h后多种基因发生了表达变化 ,涉及细胞生理多方面的功能 ,其中一些基因在癌变的早期起重要的作用 ,这为深入探讨肿瘤早期形成机制打下基础 。An inducible PTPα expression system was used to screen differential expression genes in NIH3T3 cells overexpressing PTPα, which could be used as a model for studying mechanism of tumorigenesis at early stage. After PTPα induction for 24 h, 65 differential bands were obtained, among which, there were 29 known genes, 12 known EST, 6 novel EST by bioinformatic analysis. Functions of the known genes were related to signal transduction, structural/adhesion, energy metabolism, transcription/translation, ribosome and apoptosis. The differential expression of the genes coding for G protein, TCR, Trx related protein, caveolin 1 was verified by RT PCR and Northern blot. The results showed that the expression of numerous genes was changed after the induction of PTPα for 24 h, these genes are related to many cellular physiological functions,and some of them play important roles in the early stage of tumorigenesis. The results provide basis for further exploring the molecular mechanism of tumorigenesis and these genes may well be candidate targets for gene therapy.

关 键 词：PTPα 高表达 NIH3T3细胞恶变 相关基因 筛选 鉴定 早期基因 差别表达 肿瘤形成机制 

分 类 号：R730.2[医药卫生—肿瘤]