Cloning of novel tumor metastasis-related genes from the highly metastatic human lung adenocarcinoma cell line Anip973.
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.
Liu
Fangli
F
Laboratory of Medical Genetics, Harbin Medical University, Harbin 150086, China.
Li
Yu
Y
Yu
Yang
Y
Fu
Songbin
S
Li
Pu
P
D000230:Adenocarcinoma / Q000235:genetics*; D000818:Animals; D002289:Carcinoma, Non-Small-Cell Lung / Q000235:genetics*; D003001:Cloning, Molecular; D004273:DNA, Neoplasm / Q000032:analysis*; D015972:Gene Expression Regulation, Neoplastic; D015723:Gene Library; D016147:Genes, Tumor Suppressor / Q000502:physiology*; D006801:Humans; D008175:Lung Neoplasms / Q000235:genetics*; D051379:Mice; D041681:NIH 3T3 Cells; D009362:Neoplasm Metastasis / Q000235:genetics*; D012269:Ribosomal Proteins / Q000235:genetics* / Q000378:metabolism / Q000502:physiology; D014162:Transfection; D014407:Tumor Cells, Cultured
D004273:DNA, Neoplasm; D012269:Ribosomal Proteins; C032493:ribosomal protein L17