Detection of ras oncogenes by analysis of p21 proteins in human tumor cell lines.
To detect mutationally activated ras oncogenes, we analyzed electrophoretic mobilities of ras p21 proteins utilizing the fact that many ras oncogenes produce abnormal p21 proteins that migrate at SDS/polyacrylamide gel electrophoresis as a fast-moving or slow-moving species in comparison to a normal p21 depending on the kind of mutation. Of 18 human tumor cell lines analyzed, four (SW480, SW620 and SW403 colon cancers, and SW626 ovary cancer) produced p21 belonging to the slow-moving species, suggesting a point mutation within codon 12 of a member of the three ras genes, H-, Ki- and N-ras. Subsequent DNA transfection analysis using NIH/3T3 cells as recipients identified activated Ki-ras oncogenes in the same four but not in other 14 cell lines. Thus, the analysis of p21 might serve as a rapid primary method to screen a large number of tumor materials for the presence of certain types of mutationally activated ras oncogenes.
Fujita
J
J
Institute for Cancer Research, Osaka University Medical School, Japan.
Yoshida
O
O
Ebi
Y
Y
Nakayama
H
H
Onoue
H
H
Rhim
J S
JS
Kitamura
Y
Y
D015153:Blotting, Western; D004252:DNA Mutational Analysis; D004591:Electrophoresis, Polyacrylamide Gel; D011905:Genes, ras; D006801:Humans; D015689:Oncogene Protein p21(ras); D009856:Oncogene Proteins, Viral / Q000235:genetics*; D010761:Phosphorus Radioisotopes; D014162:Transfection; D014407:Tumor Cells, Cultured
D009856:Oncogene Proteins, Viral; D010761:Phosphorus Radioisotopes; D015689:Oncogene Protein p21(ras)