甲状腺癌相关基因筛选、克隆、定位及生物学功能的研究

WEN Gebo, et al. Clinical research Institute ，The First Hospital , Nanhua University, Hengyang 421001 , China.[Abstract] Objective To clone and identify differentially expressed genes which are related to thyroid carcinoma. Methods (1) Using the technique of SSH to construct a subtractive library containing differentially expressed genes' fragments. (2) Cloning and primarily analysising those differentially expressed ESTs. (3) Homology analysis of all cDNA fragments from SSH by Blastn through GenBank .（4）Using the technique of SmartRACE to obtain the full-length sequences of new Genes Result The human Thyroid carcinoma cell (TCC) subtractive library with high subtractive efficiency was successfully constructed. Homology analysis of all 3 cDNA fragments from forward SSH by Blastn through GenBank showed high homology with some known "tumor related gene". On the other hand , 5 cDNA fragments from reverse SSH showed low homology with any known gene of human beings, it's indicated that those 5 cDNA fragments were from novel genes which specifically down-regulated expressed in Thyroid carcinoma cell. Conclusion Through using SSH, we have successfully constructed the human TCC subtractive library and therefore formed a solid foundation for identifying and full-length cloning novel genes related to thyroid carcinoma cells.

用SSH法筛选甲状腺癌EST片段，将这些片段克隆及亚克隆，测序并确定有意义的cDNA，结合人类基因组文库的匹配分析法找出长的cDNA序列。应用FISH技术确定新筛选的甲状腺癌相关基因在染色体的位置。进一步利用反义技术和转基因技术对所筛选的甲状腺癌相关基因进行生物学功能鉴定。为研究甲状腺癌发生的分子机理、基因诊断与治疗开辟新的途径。......