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TSG101, identified by screening a cancer cDNA library and soft agar assay, promotes cell proliferation in human lung cancer

Liu, Fangli, Yu, Yang, Jin, Yan, Fu, Songbin
Molecular biology reports 2010 v.37 no.6 pp. 2829-2838
Western blotting, adenocarcinoma, agar, cDNA libraries, cell proliferation, complementary DNA, genes, humans, interphase, lung neoplasms, messenger RNA, metastasis, mice, nucleotide sequences, phenotype, reverse transcriptase polymerase chain reaction, screening
Understanding the genesis and development of tumors is an essential component in cancer research. It is of interest to discover unknown genes that are responsible for cellular transformation. A cDNA library of a highly metastatic lung adenocarcinoma cell line was constructed. This library was introduced into the NIH3T3 mouse embryonic fibroblast cell line to screen for cDNAs that increase anchorage-independent colony formation in soft agar. The expression of TSG101 in lung cancer cell lines and specimens was confirmed using reverse transcription-polymerase chain reaction. The level of TSG101 protein in transfected A549 cells was determined by western blotting. Cell-cycle distribution was analyzed using a FACStar Plus flow cytometer. One of the candidate cDNAs that increases anchorage-independent colony formation was shown to correspond to the TSG101 cDNA sequence. Levels of TSG101 mRNA were higher in lung cancer cell lines and specimens compared to matched normal lung tissues. Ectopic expression of TSG101 in the A549 lung adenocarcinoma cell line increased the numbers of cells in S phase, suggesting an increased cell proliferation rate. These results indicate that TSG101 may induce the malignant phenotype of cells.