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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 ,也可为基因治疗候选靶点的选择提供依据
The established expression pattern of protein tyrosine phosphatase α (PTPα) in NIH3T3 NIH3T3 cells differentially expressed before and after induced expression of genes, is conducive to explore the mechanism of early tumor formation. The expression of PTPα was induced for 24 hours. Sixty-five differential fragments were obtained by differential display reverse transcription polymerase chain reaction (RT-PCR). The difference fragments were analyzed by bioinformatics methods and found to contain 29 known genes, 12 known ESTs, 6 unknown ESTs The results showed that they were related to signal transduction, cytoskeleton and adhesion, transcription and translation, energy metabolism, apoptosis and ribosomal proteins respectively. Among them, G protein, TCR and Trx The caveolin 1 gene was verified by RT PCR and Northern blotting. The results showed that PTPα induced expression of a variety of gene expression changes after 24 h, involving many aspects of cell physiology, some of which play an important role in the early stage of carcinogenesis, which lay the foundation for in-depth discussion of the mechanism of tumor formation in the early, but also Provide the basis for the selection of gene therapy candidate targets