Summary
Transfection of the Ha-ras oncogene into a low metastatic epithelial cell line resulted in the acquirement of significantly increased metastatic capacity. This alteration in metastatic competence of a carcinoma line in a syngeneic system seemed to be a selective change and was not affected by parameters such as tumor latency period or local tumor growth. Transfection of the selection marker vectors with normal cellular DNA or with the N-ras gene did not lead to significantly increased metastatic capacity. Analysis of metastatic variants after oncogene transfection and in vivo selection showed integration of N-ras, but not of Ha-ras oncogenes. A possible role for the Ha-ras oncogene in the initial steps of metastasis will be discussed.
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Bradley MO, Kraynak A, Storer RP, Gibbs JB (1986) Experimental metastasis in nude mice of NIH 3T3 cells containing various ras genes. Proc Natl Acad Sci USA 83:5277–5281
Brady G, Funk A, Mattern J, Schütz G, Brown R (1985) Use of gene transfer and a novel cosmid rescue strategy to isolate transforming sequences. EMBO J 4:2583–2588
Brunner G, Pohl J, Radler-Pohl A, Schirrmacher V (1988) Induction of urokinase activity and malignant phenotype in bladder carcinoma cells after transfection of the activated Ha-ras oncogene. J Cancer Res Clin Oncol (in press)
Collard JG, Schijven JF, Roos E (1987) Invasive and metastatic potential induced by ras transfection into mouse BW5147 T-lymphoma cells. Cancer Res 47:754–759
Cowell IF, Franks LM (1984) The ability of normal mouse cells to reduce the malignant potential of transformed mouse bladder epithelial cells depends on their somatic origin. Int J Cancer 33:657–667
Eccles SA, Marshall CJ, Vousden K, Purvies HP (1985) Enhanced spontaneous metastasis capacity of mammary carcinoma cells transfected with Ha-ras. In: Treatment of metastasis. Problems and prospects. Taylor and Francis, pp 385–388
Egan SE; McClarty GA, Jarolim L, Wright JA, Spiro F, Hager G, Greenberg AH (1987) Expression of the H-ras correlates with metastatic potential: Evidence for direct regulation of the meta static phenotype in 10T1/2 and NIH3T3 cells. Mol Cell Biol 7:830–837
Feinberg AP, Vogelstein B, Droller MJ, Baylin SB, Nelkin BD (1983) Mutation affecting the 12th amino acid of the c-Ha-ras oncogene product occurs infrequently in human cancer. Science 222:1175–1177
Fujita J, Srivastava SK, Kraus MH, Rhim JS, Tronick SR, Aaronson SA (1984) Frequency of molecular alterations affecting ras proto-oncogenes in human urinary tract tumors. Proc Natl Acad Sci 82:3849–3853
Furth ME, Davis LJ, Fleudelys B, Scolnick EM (1982) Monoclonal antibody to the p21 products of the transforming gene of Harvey murine sarcoma virus and the cellular ras gene family. J Virol 43:294–304
Gallick GE, Kurzrock R, Kloetzer WS, Arlinghaus RB, Gutterman JU (1985) Expression of p21ras in fresh primary and metastatic human colorectal tumors. Proc Nate Acad Sci USA 82:1795–1799
Glover MD (1985) DNA cloning, vol 2, a practical approach. IRL Press Limited
Gorman CM, Leslie FM, Howard BH (1982) Recombinant genomes which express chloramphenicol acetyltransferase in mammalian cells. Mol Cell Biol 2:1044–1051
Greig G, Kostler TP, Trainer DL, Corwin SP, Miles L, Kline T, Sweet R, Yokoyama S, Poste G (1985) Tumorigenic and metastatic properties of “normal” and ras-transfected NIH/3T3 cells. Proc Natl Acad Sci USA 82:3698–3701
Kerbel R, Waghorne C, Man MS, Elliott B, Breitman ML (1987) Alteration of the tumorigenic and metastatic properties of neoplastic cells is associated with the process of calcium phosphate mediated DNA transfection. Proc Natl Acad Sci USA 84:1263–1267
Land H, Parada LF, Weinberg RA (1983) Tumorigenic conversion of primary embryo fibroblasts requires at least two cooperating oncogenes. Nature 304:596–602
Maniatis T, Fritsch EF, Sambrock J (1982) Molecular cloning. Cold Spring Harbor Laboratory
Muschel RJ, Williams JE, Lowy DR, Liotta LA (1985) Harvey ras induction of metastatic potential depends upon oncogene activation and the type of recipient cell. Am J Pathol 121:1–8
Parada LF, Land H, Weinberg RA, Wolf D, Rotter V (1984) Cooperation between gene encoding p53 tumor antigen and ras in cellular transformation. Nature 312:649–651
Pohl J, Radler-Pohl A, Heicappell R, Schirrmacher V (1987) Oncegene expression in cell lines differing in metastatic capacity. Clin Exp Met 6:201–211
Pozzatti R, Muschel R, Williams J, Padmanabhan R, Howard B, Liotta L, Khoury G (1986) Primary rat embryo cells transformed by one or two oncogenes show different metastatic potentials. Science 232:223–227
Radler-Pohl A, Pohl J, Schirrmacher V (1988) Selective enhancement of metastatic capacity in mouse bladder carcinoma cells after transfection with DNA from human colon liver metastases. Int J Cancer (in press)
Reddy EP, Reynolds RK, Santos E, Barbacid M (1982) A point mutation is responsible for the aquisition of transforming properties by the T24 human bladder carcinoma oncogene. Nature 300:149–152
Rigby PW, Dieckmann M, Rhodes C, Berg P (1977) Labeling deoxy-ribonucleic acid to high specific activity in vitro by nick translation with DNA polymerase I. Proc Natl Acad Sci USA 113:237–251
Scangos G, Ruddle FH (1981) Mechanism and application of DNA-mediated gene transfer in mammalian cells — a review. Gene 14:1–10
Sistonen L, Keski-Oja J, Ulmanen I, Hölttä E, Wilkren B-J, Alitalo K (1987) Dose effects of transfected c-Ha-rasVal12 oneogene in transformed cell clones. Exp Cell Res 168:518–530
Southern PJ, Berg P (1982) Transformation of mammalian cells to antibiotic resistence with a bacterial gene under control of the SV40 early region promoter. J Mol Appl Genet 1:327–341
Stenman G, Delorme EO, Lau CC, Sager R (1987) Transfection with plasmid pSVgptEJ induces chromosome rearrangements in CHEF cells. Proc Natl Acad Sci USA 84:184–188
Summerhayes IC, Malone P, Visvanathan K (1986) Altered growth properties and cell surface changes in ras transformed mouse bladder epithelium. Int J Cancer 37:233–240
Temin HM (1979) Malignant transformation of cells by viruses. Perspect Biol Med 14:11–26
Thorgeirsson UP, Turpeenniemi-Hujanen T, Williams JE, Westin EH, Heihman CA, Talmadge JE, Liotta LA (1985) NIH/3T3 cells transfected with human tumor DNA containing activated ras oncogenes express the metastatic phenotype in nude mice. Mol Cell Biol 5:259–262
Varani J, Fligiel SEG, Wilson B (1986) Motility of rasH oncogene transformed NIH-3T3 cells. Inv Met 6:335–346
Vousden KH, Eccles SA, Purvies P, Marshall CJ (1986) Enhanced spontaneous metastasis of mouse carcinoma cells transfected with an activated c-Ha-ras-1 gene. Int J Cancer 37:425–433
Yokota J, Tsunetsugu-Yokota Y, Battifora H, Lefevre C, Cline MJ (1986) Alterations of myc, myb, and rasHa proto-oncogenes in cancers are frequent and show clinical correlation. Science 231:261–265
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Pohl, J., Radler-Pohl, A., Franks, L.M. et al. Analysis of metastatic competence of mouse bladder carcinoma cells after transfection with activated Ha-ras or N-ras oncogenes. J Cancer Res Clin Oncol 114, 373–379 (1988). https://doi.org/10.1007/BF02128181
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DOI: https://doi.org/10.1007/BF02128181