Ki-ras activation and expression in transformed mouse lung cell lines

✍ Scribed by Y. H. Laurie Pan; Edwin O. Nuzum; Laurie A. Hanson; David G. Beer

• Publisher: John Wiley and Sons
• Year: 1990
• Tongue: English
• Weight: 754 KB
• Volume: 3
• Category: Article
• ISSN: 0899-1987
• DOI: 10.1002/mc.2940030508

✦ Synopsis

Abstract

The Ki‐ras proto‐oncogene is activated by specific point mutations and is the transforming gene often identified in rodent and human lung tumors. An in vitro model to aid in the study of the consequences of Ki‐ras activation and expression in mouse lung is needed. Accordingly, we have examined cell lines derived from chemically induced mouse lung tumors as well as spontaneous transformants of untreated mouse lung epithelial cells. The specific Ki‐ras‐activating gene mutations and the level of mRNA expression were examined for each cell line. Polymerase chain reaction and oligonucleotide hybridization were used to demonstrate that five of seven transformed lung cell lines contain codon 61 Ki‐ras‐activating mutations, resulting in an arginine substitution for wild‐type glutamine. One transformed line contained this activating mutation and had also lost, or contained an altered, wild‐type codon 61 Ki‐ras allele. No codon 12 Ki‐ras mutations were observed. Two transformed and two nontransformed epithelial lung cell lines contained only the wild‐type codon 12 and 61 Ki‐ras alleles. Northern blot analysis demonstrated that the Ki‐ras mRNA was present in all the cell lines and was overexpressed in some, but not all, of the transformed lung cell lines. Those transformed lines with the highest levels of Ki‐ras mRNA also expressed more H4‐histone mRNA, suggesting that these cells have a greater proliferation rate. The level of Ki‐ras mRNA increased during the proliferation of the nontransformed lung cells but then decreased upon reaching confluency. In contrast, the level of Ki‐ras mRNA in the transformed lung cells was high during both growth and confluency, suggesting a potential defect in the regulation of Ki‐ras in these cells. These lung cell lines will help provide a better understanding of the regulation of both the Ki‐ras proto‐oncogene and oncogene in the lung.