Abstract
Human breast cancer cell lines which grow in athymic (nude) mice provide a model of tumor cell growth and metastasis. Marking transplanted tumor cell populations with retroviral vectors provides a means of studying the dynamics of tumor cell growthin vivo. We evaluated three human breast cancer cell lines, MDA-MB-435, MDA-MB-231 and MCF-7, and found the cells were highly susceptible to retroviral gene transfer after a single 2-h exposure (90.9%, 62.7% and 72.3%, respectively). MDA-MB-435 cells (5×105) marked with a retroviral vector containing theβ-galactosidase gene (approximately 104 uniquely marked clones) were injected into the mammary fat pad of athymic mice to study clonal dominance. Primary tumors resected 10 weeks after injection expressedβ-galactosidase, demonstrating persistent vector expressionin vivo. Southern blot analysis did not reveal clonal dominance in the primary tumors of the five mice studied. In contrast, pulmonary metastases in each animal were monoclonal or biclonal. These results demonstrate clonal dominance in pulmonary metastases but not primary tumors of retrovirally marked MDA-MB-435 cells. Our findings suggest that this model may also be used to introduce retroviral vectors expressing oncogenes, and anti-sense oncogenes, to determine their effect on tumor cell proliferation and metastasisin vivo.
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References
Shafie SM and Liotta LA, 1980, Formation of metastasis by human breast carcinoma cells (MCF-7) in nude mice.Cancer Letters,11, 81–87.
Ozzello L and Sorbat M, 1980, Behavior of tumors produced by transplantation of human mammary cell lines in athymic nude mice.European Journal of Cancer,16, 553–559.
Busuttil A, O'Conor GT, Foster ME, Gurtsevitch V, Morten JEN and Steel CM, 1986, The gross pathology and histology features of tumors produced by inoculation of human cell lines into immunodeprived mice.Journal of Pathology,148, 293–300.
Price JE, Polyzos A, Zhang RD and Daniels LM, 1990, Tumorigenicity and metastasis of human breast carcinoma lines in nude mice.Cancer Research,50, 717–721.
Williams DA, Lemischka IR, Nathan DG and Mulligan RC, 1984, Introduction of new genetic material into pluripotent hematopoietic stem cells of the mouse.Nature,310, 476–480.
Waghorne C, Thomas M, Lagarde A, Kerbel RS and Breitman ML, 1988, Genetic evidence for progressive selection and overgrowth of primary tumors by metastatic cell subpopulations.Cancer Research,48, 6109–6114.
Rosenberg SA, Aebersold P, Cornetta K, Kasid A, Morgan RA, Karson E, Lotze MT, Yang YC, Topalian S, Moen RC, Culver K, Blaese RM and Anderson WF, 1990, Gene transfer into humans: immunotherapy of patients with advanced melanoma using tumor infiltrating lymphocytes modified by retroviral gene transduction.New England Journal of Medicine,323, 570–578.
Korczak B, Robson IB, Lamarche C, Bernstein A and Kerbel RS, 1988, Genetic tagging of tumor cells with retrovirus vectors: clonal analysis of tumor growth and metastasisin vivo.Molecular and Cellular Biology,8, 3143–3149.
Price JE, Bell C and Frost P, 1990, The use of a genotypic marker to demonstrate clonal dominance during the growth and metastasis of a human breast carcinoma in nude mice.International Journal of Cancer,45, 968–971.
Miller AD and Rosman GJ, 1989, Improved retroviral vectors for gene transfer and expression.BioTechniques,7, 980–990.
Miller AD and Buttimore C, 1986, Redesign of retrovirus packaging cell lines to avoid recombination leading to helper virus production.Molecular and Cellular Biology,6, 2895–2902.
Kantoff PW, Kohn DB, Mitsuya H, Armentano D, Sieberg M, Zwiebel JA, Eglitis MA, McLachlin JR, Wiginton DA, Hutton JJ, Horowitz SD, Gilboa E, Blaese RM and Anderson WF, 1986, Correction of adenosine deaminase deficiency in cultured human T and B cells by retrovirus-mediated gene transfer.Proceeding of the National Academy of Sciences USA,83, 6563–6567.
Cornetta K, Moen RC, Culver K, Morgan RA, McLachlin JR, Sturm S, Selegue JE, London WT, Blaese RM and Anderson WF, 1990, Amphotropic murine leukemia retrovirus is not an acute pathogen for primates.Human Gene Therapy,1, 14–30.
Lim K and Chae CB, 1989, A simple assay for DNA transfection by incubation of the cells in culture dishes with substrates forβ-galactosidase.BioTechniques,7, 576–579.
Southern EM, 1975, Detection of specific sequences among DNA fragments separated by gel electrophoresis.Journal of Molecular Biology,98, 503–517.
Reiss B, Sprengel R, Will H and Schaller H, 1984, A new sensitive method for qualitative and quantitative assay for neomycin phosphotransferase in crude cell extracts.Gene,30, 211–226.
Brunner N, Thompson EW, Spang-Thomsen M, Rygaard J, Dano K and Zwiebel JA, 1992, lacZ Transduced human breast cancer xenografts asin vivo model for the study of invasion and metastasis.European Journal of Cancer,28A, 1989–1995.
Palmer TD, Rosman GJ, Osborne WR and Miller AD, 1991, Genetically modified skin fibroblasts persist long after transplantation but gradually inactivate introduced genes.Proceedings of the National Academy of Sciences USA,88, 1330–1334.
Emerman M and Temin HM, 1984, Genes with promoters in retrovirus vectors can be independently suppressed by an epigenetic mechanism.Cell,39, 459–467.
Emerman M and Temin HM, 1984, High-frequency deletion in recovered retrovirus vectors containing exogenous DNA with promoters.Journal of Virology,50, 42–49.
Kerbel RS, Waghorne C, Man MS, Elliott B and Breitman ML, 1987, Alteration of the tumorigenic and metastatic properties of neoplastic cells is associated with the process of calcium phosphate-mediated DNA transfection.Proceedings of the National Academy of Sciences USA,84, 1263–1267.
Heppner GH, Miller BE and Miller FR, 1983, Tumor subpopulation interactions in neoplasms.Biochimica et Biophysica Acta,695, 215–226.
Nowell PC, 1976, The clonal evolution of tumor cell populations.Science,194, 23–28.
Rubin H, 1984, Early origin and pervasiveness of cellular heterogeneity in some malignant transformations.Proceedings of the National Academy of Sciences USA,81, 5121–5125.
Tisty TD, Margolin BH and Lum K, 1989, Differences in the rates of gene amplification in nontumorigenic and tumorigenic cell lines as measured by Luria-Delbruck fluctuation analysis.Proceedings of the National Academy of Sciences USA,86, 9441–9445.
Moffett BF, Baban D, Bao L and Tarin D, 1992, Fate of clonal lineages during neoplasia and metastasis studied with an incorporated genetic marker.Cancer Research,52, 1737–1743.
Meyer JS and Wittliff JL, 1991, Regional heterogeneity in breast carcinoma: thymidine labeling index, steroid hormone receptors, DNA ploidy.International Journal of Cancer,47, 213–220.
Van Diest PJ, Fleege JC, Matze-Cok E and Baak JPA, 1992, Intrapatient variation between breast cancer axillary lymph node metastases using quantifiable features.Histopathology,21, 257–262.
Kallionemi O-P, Kallionemi A and Kurisu W, 1992, ERB-B2 amplification in breast cancer analyzed by fluorescencein situ hybridization.Proceedings of the National Academy of Sciences USA,89, 5321–5325.
Chen L, Kurisu W, Lijung B-M, Goldman ES, Moore D and Smith HS, 1992, Heterogeneity for allelic loss in human breast cancer.Journal of the National Cancer Institute,84, 506–510.
Hill RP, Chambers AF, Ling V and Harris JF, 1984, Dynamic heterogeneity: rapid generation of metastatic variants in mouse B16 melanoma cells.Science,224, 998–1001.
Harris JF, Chambers AF, Hill RP and Ling V, 1982, Metastatic variants are generated spontaneously at a high rate in mouse KHT tumor.Proceedings of the National Academy of Sciences USA,79, 5547–5551.
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Cornetta, K., Moore, A., Johannessohn, M. et al. Clonal dominance detected in metastases but not primary tumors of retrovirally marked human breast carcinoma injected into nude mice. Clin Exp Metast 12, 3–12 (1994). https://doi.org/10.1007/BF01784328
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DOI: https://doi.org/10.1007/BF01784328