Abstract
The transformation frequency of mouse BALB/3T3 cells was significantly enhanced after transfection with an activated ras oncogene (v‐Ha‐ras) followed by treatment with a tumor promoter, 12‐O‐tetradecanoyl‐phorbol‐13‐acetate (TPA), suggesting that the ras oncogene acted as an initiator in two‐stage carcinogenesis. A cell clone (Bhas42) containing the ras oncogene was isolated from the ras‐transfected BALB/3T3 cells. Bhas42 cells were flat and showed contact inhibition, but the addition of TPA to quiescent Bhas42 cultures resulted in a dramatic change of cell morphology to spindle shape, doubling of the cell population, and increased DNA synthesis.
Keywords: Transformation, ras oncogene, TPA, BALB/3T3
Full Text
The Full Text of this article is available as a PDF (643.5 KB).
References
- 1).Boutwell , R. K.The function and mechanism of promoters of carcinogenesis . CRC Crit. Rev. Toxicol. , 2 , 419 – 431 ( 1974. ). [DOI] [PubMed] [Google Scholar]
- 2).Mondal , S.Brankow , D. W. and Heidelberger , C.Two‐stage chemical carcinogenesis in cultures of C3H/10T1/2 cells . Cancer Res. , 36 , 2254 – 2260 ( 1976. ). [PubMed] [Google Scholar]
- 3).Wynder , E. L. , Hoffman , D. , McCoy , G. D. , Cohen , L. A. and Reddy , B. S.Tumor promotion and cocarcinogenesis as related to man and his environment . In “ Carcinogenesis ,” ed. Slaga T. J. , Sivak A. and Boutwell R. K. , Vol. 2 , pp . 59 – 77 ( 1978. ). Raven Press; , New York . [Google Scholar]
- 4).Little , J. B.Cellular mechanisms of oncogenic transformation in vitro . In “ Transformation Assay of Established Cell Lines: Mechanisms and Application ,” ed Kakunaga T. and Yamasaki H. , IARC Scientific Publications , Vol. 67 , pp . 9 – 30 ( 1985. ). IARC; , Lyon . [PubMed] [Google Scholar]
- 5).Berenblum , I.The cocarcinogenic action of croton resin . Cancer Res. , 1 , 44 – 48 ( 1940. ). [Google Scholar]
- 6).Brown , K. , Quintanilla , M. , Ramsden , M. , Ker , I. B. , Young , S. and Balmain , A.v‐ras genes from Harvey and BALEB murine sarcoma viruses can act as initiators of two‐stage mouse skin carcinogenesis . Cell , 46 , 447 – 456 ( 1986. ). [DOI] [PubMed] [Google Scholar]
- 7).Boutwell , R. K.Biochemical mechanism of tumor promotion . In “ Carcinogenesis ,” ed. Slaga T. J. , Sivak A. and Boutwell R. K. , Vol. 2 , pp . 49 – 58 ( 1978. ). Raven Press; , New York . [Google Scholar]
- 8).Sasaki , K. , Chida , K. , Hashiba , H. , Kamata , T. , Abe , E. , Suda , T. and Kuroki , T.Enhancement by lα,25‐dihydroxyvitamin D3 of chemically induced transformation of BALEB/3T3 cells without induction of ornithine decarboxylase or activation of protein kinase C . Cancer Res. , 46 , 604 – 610 ( 1986. ). [PubMed] [Google Scholar]
- 9).Hsiao , W. L‐W. , Gattoni‐Celli , S. and Weinstein , I. B.Oncogene induced transformation of C3H10T1/2 cells is enhanced by tumour promoters . Science , 226 , 552 – 553 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 10).Dotto , G. P. , Parada , L. F. and Weinberg , R. A.Specific growth response of ras‐transformed embryo fibroblasts to tumour promoters . Nature , 318 , 472 – 475 ( 1985. ). [DOI] [PubMed] [Google Scholar]
- 11).Kakunaga , T. and Crow , J. D.Cell variants showing differential susceptibility to ultraviolet light‐induced transformation . Science , 209 , 505 – 507 ( 1980. ). [DOI] [PubMed] [Google Scholar]
- 12).Ellis , R. W. , De Feo , D. , Maryak , J. M. , Young , H. A.Shih , T. Y.Chang , E. H. , Lowy , D. R. and Scolnick , E. M.Dual evolutionary origin for the rat genetic sequences of Harvey murine sarcoma virus . J. Virol. , 36 , 408 – 420 ( 1980. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 13).Bazaral , M. and Helinski , D. R.Circular DNA of colicinogenic factors. E1, E2, and E3 from Escherichia coli . J. Mol. Biol. , 36 , 185 – 194 ( 1968. ). [DOI] [PubMed] [Google Scholar]
- 14).Mizusawa , H. , Tanaka , S. , Kobayashi , M. and Koike , K.Alkali‐labile colicinogenic factor E1 DNA molecules formed in the presence of N‐methyl‐N' ‐nitro‐N‐nitroso‐guanidine . Biochem. Biophys. Res. Commun. , 74 , 570 – 576 ( 1987. ). [DOI] [PubMed] [Google Scholar]
- 15).Wigler , M. , Pellicer , A. , Silverstein , S. , Axel , R. , Urlaub , G. and Chasin , L.DNA‐mediated transfer of the adenine phospho‐ribosyltransferase locus into mammalian cells . Proc. Natl. Acad. Sci. USA , 76 , 1373 – 1376 ( 1979. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 16).Lewis , W. H. , Srinivasan , P. R. , Stokoe , N. and Siminovitch , L.Parameters governing the transfer of the genes for thymidine kinase and dehydrofolate reductase into mouse cells using metaphase chromosome or DNA . Somatic Cell. Genet. , 6 , 333 – 347 ( 1980. ). [DOI] [PubMed] [Google Scholar]
- 17).Muller , R. and Wagner , E. F.Differentiation of F9 teratocarcinoma stem cells after transfer of c‐fos proto‐oncogenes . Nature , 311 , 438 – 442 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 18).Parnes , J. R. , Velan , B. , Felsenfeld , A. , Ramanathan , L. , Ferrini , U. , Appella , E. and Seidman , J. G.Mouse B2 microglobulin cDNA clones: a screening procedure for cDNA clones corresponding to rare mRNAs . Proc. Natl. Acad. Sci. USA , 78 , 2253 – 2257 ( 1981. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 19).Maniatis , T. , Jeffrey , A. and Kleid , D. G.Nucleotide sequence of the rightward operator of phage lambda . Proc. Natl. Acad. Sci. USA , 72 , 1184 – 1188 ( 1975. ). [DOI] [PMC free article] [PubMed] [Google Scholar]
- 20).Southern , E.Gel electrophoresis of restriction fragments . Methods Enzymol. , 68 , 152 – 176 ( 1979. ). [DOI] [PubMed] [Google Scholar]
- 21).Jones , C. A. , Callaham , M. F. and Huberman , E.Enhancement of chemical carcinogen‐induced cell transformation in hamster embryo cells by lα,25‐dihydroxy‐cholecalciferol, the biological active vitamin D3 . Carci-nogenesis , 5 , 1155 – 1159 ( 1984. ). [DOI] [PubMed] [Google Scholar]
- 22).Umeda , M. , Tanaka , K. and Ono , T.Effect of insulin on the transformation of BALB/3T3 cells by X‐ray irradiation . Gann , 74 , 864 – 869 ( 1983. ). [PubMed] [Google Scholar]