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Neoplastic transformation of a human bronchial epithelial cell line by a recombinant retrovirus encoding viral harvey ras
Author(s) -
Amstad Paul,
Reddel Roger R.,
Pfeifer Andrea,
MalanShibley Louise,
Mark George E.,
Harris Curtis C.
Publication year - 1988
Publication title -
molecular carcinogenesis
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.254
H-Index - 97
eISSN - 1098-2744
pISSN - 0899-1987
DOI - 10.1002/mc.2940010303
Subject(s) - biology , retrovirus , cell culture , carcinogenesis , neoplastic transformation , virology , cancer research , oncogene , immortalised cell line , cell , microbiology and biotechnology , transfection , virus , gene , cell cycle , genetics
Activated ras oncogenes have previously been implicated in the pathogenesis of human lung carcinomas. A v‐Ha‐ ras ‐containing retrovirus, Zip‐ras, was generated by inserting the coding region of the v‐Ha‐ras oncogene into the Zip‐NeoSV(X) [Cepko et al., Cell 37:1053–1062, 1984] retroviral vector. Amphotrophic Zip‐ras retrovirus was used to infect an SV40 large T antigen‐positive immortalized cell line, BEAS‐2B, derived from normal bronchial epithelial cells, the predominant progenitor cells of human lung carcinomas. Zip‐ras—infected BEAS‐2B cells selected for G418 resistance formed anaplastic carcinomas in 12 of 15 athymic nude mice (latency 3 wk), whereas Zip‐NeoSV(X)—infected BEAS‐2B control cultures inoculated into 12 nude mice formed no tumors after a minimum of 7 mo. Tumor cell lines were established and demonstrated to be of human epithelial origin and to express v‐Ha‐ ras p21 protein. A common feature of the tumor cell lines was an increase in ploidy. The increased efficiency of neoplastic transformation by v‐Ha‐ras of cell lines as compared with our previous results with normal bronchial epithelial cells [Yoakum et al., Science 227:1174–1179, 1985] is consistent with the hypothesis that the “immortalization” step is rate‐limiting in in vitro human epithelial cell carcinogenesis.