RAB5A 基因高表达在肺腺癌转移形成中的作用及其机理

In this study, in order to investigate the function and biological significance of RAB5A gene in the metastasis of human neoplasma, we constructed RAB5A eukaryotic expression plasmid and antisense RNA of RAB5A with pcDNA3.1(-) and pcDNA3 eukaryotic expression vectors which were sequently transfected into human lung adenocarcinoma cell lines. Furthermore, we carried out tumorigenesis and tumor-suppression experiments in the nude mice and in vitro. Our results indicate that the high expression level of RAB5A has some cellular genetic basis, and can influence the cytoskeletal structure, causing significant changes in the ultrastructure on the cell surface, and in the cell adhesion as well as cytotaxis relevant to cancer metastasis. Meanwhile the antisense molecule can lead to an apparent change of cell cycle and a lot of cells were retained at G0-G1 phase, which has never been reported in the previous studies. According to the above findings, we applied for and obtained the support of the project of international cooperation and exchange. Our further investigation indicated that RAB5A exerts an definit influence on the cell cycle through some regulatory pathway. In conclusion, all the results we got lead to a better understanding of the cellular biological functions of RAB5A known as a signal transduction regulator in protein endocytosis and their molecular mechanisms in the formation of metastatic phenotype in human cancer.

通过mRNA差异展示技术对两株细胞来源相同但转移能力不同的人肺腺癌细胞系AGZY-83a和Anip973的分离筛选表明RAB5A基因在高转移细胞系中高表达。本研究拟用真核表达载体构建RAB5A的正义表达载体和反义RNA，采用基因转移技术，通过体外和裸鼠体内成瘤或抑瘤试验，确认RAB5A基因在肿瘤转移中作用和生物学意义。并cDNA RDA采用方法分析作用机怼