Transformation of NIH3T3 cells by transfection with UV-irradiated human c-Ha-ras-1 proto-oncogene DNA.
Our previous studies have shown that human skin cancers occurring on sun-exposed body sites frequently contain G----T mutations at the second position of Ha-ras codon 12. In this study, we investigated whether the c-Ha-ras-1 proto-oncogene could be activated by in vitro UV-irradiation of pEC plasmid DNA, which contains the 6.6 kb BamHI fragment of the human c-Ha-ras-1 proto-oncogene. Focus formation and nude mouse tumorigenicity assays showed that UV-irradiated pEC DNA induced morphologic and tumorigenic transformation of NIH3T3 cells in multiple cycles of transfection, whereas unirradiated pEC DNA did not. DNAs from secondary cycle foci and tertiary cycle tumors were analyzed for mutations in Ha-ras codons 12 and 61 using the polymerase chain reaction and synthetic oligonucleotide probes. Eleven of 11 secondary cycle foci analyzed possessed a G----T mutation at the second position of Ha-ras codon 12. However, the nude mouse tumors exhibited a G----A mutation at position 1 of the Ha-ras codon 12. These results suggest that in vitro UV irradiation of the c-Ha-ras-1 proto-oncogene DNA can induce mutations that are similar to those found in human skin cancers that originated on sun-exposed body sites.
Pierceall
W E
WE
Department of Immunology, University of Texas M.D. Anderson Cancer Center, Houston 77030.
Ananthaswamy
H N
HN
D000818:Animals; D001483:Base Sequence; D002471:Cell Transformation, Neoplastic / Q000235:genetics*; D015972:Gene Expression Regulation, Neoplastic / Q000528:radiation effects*; D011905:Genes, ras / Q000502:physiology* / Q000528:radiation effects; D006801:Humans; D015151:Immunoblotting; D051379:Mice; D008815:Mice, Inbred Strains / Q000235:genetics*; D008969:Molecular Sequence Data; D015345:Oligonucleotide Probes / Q000235:genetics; D016133:Polymerase Chain Reaction; D000090063:Proto-Oncogene Mas; D014162:Transfection; D014466:Ultraviolet Rays / Q000009:adverse effects*
C000715470:MAS1 protein, human; D015345:Oligonucleotide Probes; D000090063:Proto-Oncogene Mas