【作者】 雷薇；

【导师】 吴旻；

【作者基本信息】 中国协和医科大学 ， 细胞生物学， 1995， 博士

【摘要】 目前认为，恶性肿瘤是一类因多个基因改变而影响细胞正常生长和分化调节的疾病。以该理论为基础的肿瘤分化治疗，目的不是单纯地杀死肿瘤细胞，而是要让那些具有分化潜能的肿瘤细胞向成熟的分化表型发展。在过去的十年中，本实验室一直在探索一条分离肿瘤促分化基因的新途径，即用分化诱导剂—全反式维甲酸诱导某些肿瘤细胞系分化，再利用cDNA文库的递减杂交策略分离出那些在分化过程中特异表达的基因，并进一步研究它们作为治疗肿瘤的目的基因的可能性。至今，已经从食管癌细胞中得到一个具有很强的肿瘤抑制功效的新基因：RATS1基因。 在上述理论及实践的指导下，本室黎伯铨等运用相同的策略，建立了维甲酸诱导前后的肺腺癌细胞系GLC-82的cDNA文库，经6轮递减杂交及初步筛选，得到124个维甲酸激活的cDNA克隆。本研究的目的即是从这些cDNA克隆中筛选出那些真正为维甲酸特异激活表达的、具有促分化功能的新基因，探讨它们与肺癌发生、发展的关系，深化维甲酸的作用机理，为克隆出这些基因以至用于肺癌的基因治疗提供基础。 本文运用限制酶切分析cDNA文库的Southern杂交，更多还原

【Abstract】 It’s known that the multistep carcinogenic process is often characterized by loss of normal control over proliferation and aberrant patterns of difFerentiation. The principle of tumor differentiation therapy has been developed on the basis of these facts. It doesn’t attempt to kill tumor cells, but focuses on the suppression of cell proliferation and the induction of terminal differentiation. During the last decade, our lab has been developing a novel strategy for isolating differentiation-inducing genes from human cancers: terminal differentiation of human cancer cell lines is induced by all-trans-retinoic acid(RA), then repeated subtraction hybridization between cDNA libraries constructed from cells before and after RA-treatrnent is used’ to isolate genes specifically activated by RA. The biological effects of these genes on parental cells are then tested in vitro. So far, a novel gene—RATS1 has been isolated from esophageal cancer cell line which possesses strong proliferation-suppressing and differentiation-inducing effect.With similar strategy, our lab also attempted to isolate those differentiation-inducing genes from lung cancer cells: three cDNA libraries were constructed, one was from human lung adenocarcinoma cell line GLC-82, named Lib.GLC-82; the other were from GLC-82 cells treated with RA for 1 and 4 days, named Lib.GLC-82.RAl and Lib.GLC-82.RA4. After the 6th round of subtraction hybridization, 124 differential cDNA clones were obtained. This study is concerned on identifying those novel genes, investigating their correlation with carcinogenesis of lung cancer, dissecting the mechanism of RA’s action, and evaluate their biological functions in their parental cells GLC-82.更多还原