cDNA表达文库转染NIH3T3-从高转移人肺腺癌细胞系Anip973中克隆新肿瘤转移相关基因.pdfVIP

Journal of Genetics and Genomics (Formerly Acta Genetica Sinica) March 2007, 34(3): 189-195 Research Article Cloning of Novel Tumor Metastasis-Related Genes from the Highly Metastatic Human Lung Adenocarcinoma Cell Line Anip973 1 2 1 1 1,① Fangli Liu , Yu Li , Yang Yu , Songbin Fu , Pu Li 1. Laboratory of Medical Genetics, Harbin Medical University, Harbin 150086, China; 2. Department of Life Science and Engineering, Harbin Institute of Technology, Harbin 150001, China Abstract: A cDNA library was successfully constructed from Anip973, a human lung adenocarcinoma cell line with high metastatic potential. NIH3T3 cells were stably transfected using this cDNA library and screened for morphological changes in a soft agar assay. Genomic DNA was isolated from putative clones and the integrated sequence was retrieved by PCR and sequencing. Three known genes, ribosomal protein L23, hypothetical protein FLJ22104, and serine protease inhibitor, kazal type 6 and a number of 5′-terminally truncated sequences were identified. Furthermore, cells transfected with ribosomal protein L23 was highly invasive compared with the empty vector as control (P 0.02). These results indicate that the expression cloning of cDNA libraries in NIH3T3 cells and subsequent screening for loss of contact inhibition in soft agar is a viable tool for identifying tumor-related genes and ribosomal protein L23 gene plays a role in cell movement and metastasis. Keywords: cDNA expression library; soft agar; oncogene; metastasis; Homo sapiens ribosomal protein L23 Research of tumor metastasis is an essential libraries can be a powerful tool for the discovery of component of the overall ca