Transformation of mouse skin endothelial cells in vivo by direct application of plasmid DNA encoding the human T24 H-ras oncogene.
Plasmid DNA containing the human T24 H-ras oncogene, with or without viral transcriptional enhancer sequences, was applied to scarified mouse skin, followed by multiple treatments with the tumour promoter 12-O-tetradecanoyl-phorbol-13-acetate. This resulted in the formation of vasoformative tumours histologically characterized as lymphangiosarcomas. All of the animals treated developed cystic fluid-filled swellings. Polymerase chain reaction analysis revealed the presence of human H-ras sequences within the cystic fluid from 3 out of 4 swellings. An endothelial cell line established from the cystic fluid removed from one of these swellings was found to contain human H-ras sequences and to express the mutant human p21ras. Injection of the cell line into nude mice, or adult syngeneic mice, resulted in the formation of aggressive angiosarcomas. Further experiments showed that 12-O-tetradecanol-phorbol-13-acetate promotion is not required for tumour formation and would appear to reduce the yield of tumours. These results indicate that a single application of the human H-ras oncogene is sufficient to induce endothelial cell transformation in vivo, even in the absence of any further promotional stimulus.
Burns
P A
PA
CRC Beatson Laboratories, Beatson Institute for Cancer Research, Glasgow, UK.
Jack
A
A
Neilson
F
F
Haddow
S
S
Balmain
A
A
D000818:Animals; D001483:Base Sequence; D015139:Blotting, Southern; D002471:Cell Transformation, Neoplastic; D003001:Cloning, Molecular / Q000379:methods*; D004591:Electrophoresis, Polyacrylamide Gel; D011905:Genes, ras; D006801:Humans; D051379:Mice; D008815:Mice, Inbred Strains; D008969:Molecular Sequence Data; D009374:Neoplasms, Experimental / Q000235:genetics*; D010957:Plasmids; D016133:Polymerase Chain Reaction; D011233:Precipitin Tests; D012878:Skin Neoplasms / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate
D013755:Tetradecanoylphorbol Acetate