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Characterization of brain and bone-metastasizing clones selected from an ethylnitrosourea-induced rat mammary carcinoma

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Abstract

An animal model for breast cancer, brain and bone metastasis was developed using ENU1564, a cell line established from a metastatic mammary adenocarcinoma induced by N-ethyl-N-nitrosourea in a female Berlin-Druckrey IV rat. The original tumor isolate (designated FP1) spontaneously metastasizes to regional lymph nodes and lung following orthotopic inoculation into mammary fat pad (mfp) and metastasizes widely following left cardiac ventricle (LV) inoculation. From FP1, two sublines were selected from brain metastases (designated Br7-C5) or from slowly growing coloniesin vitro (FP2-A11), then cloned and compared in assays of spontaneous and experimental metastasis. After inoculation of 105 cells into mfp, Br7-C5 formed large tumors at the inoculation site (9.4±3.3 g) and spontaneously metastasized to lung and lymph node by 55 days post-inoculation (dpi). In contrast, FP2-A11 produced much smaller tumors within mfp (0.6±0.3 g) and failed to metastasize by 55 dpi. Rats inoculated via the LV with 104 Br7-C5 cells developed signs of weight loss, head tilt, and dyspnea by 24±1.4 dpi with consistent colonization of brain, bone, lung, heart, kidney, and stomach. Rats inoculated similarly with FP2-A11 showed no signs until 53±12.3 dpi, when all developed rear limb paresis. There was significant colonization of only brain and bone, with only minor lung involvement. These ENU1564 sublines thus differ in their apparent rates of tumor growth and lesion developmentin vivo, their capacity to metastasize from orthotopic implantation sites, and in the spectrum of tissues colonized in experimental metastasis assays. Both clones provide reproducible models of breast cancer metastasis in syngeneic hosts, particularly to brain and bone.

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References

  1. Boring CC, Squires TS and Tong T, 1993, Cancer statistics.Ca Cancer J Clin,43, 7–26.

    PubMed  Google Scholar 

  2. Patanaphan V, Salazar OM and Risco R, 1988, Breast cancer: metastatic patterns and their prognosis.Southern Med J,81, 1109–12.

    Google Scholar 

  3. Cho SY and Choi HY, 1980, Causes of death and metastatic patterns in patients with mammary cancer.Am J Clin Pathol,73, 232–4.

    PubMed  Google Scholar 

  4. Kamby C, 1990, The pattern of metastases in human breast cancer: methodological aspects and influence of prognostic factors.Cancer Treat Rev,17, 37–61.

    Google Scholar 

  5. Namer M, 1991, Clinical consequences of osteolytic bone metastases.Bone,12 (Suppl. 1), S7.

    PubMed  Google Scholar 

  6. Greenberg EJ, 1991, The treatment of metastatic breast cancer.Ca Cancer J Clin,41, 242–56.

    PubMed  Google Scholar 

  7. Russell DS and Rubenstein LJ, 1989, Secondary Tumours. In: Russell DS and Rubenstein LJ, eds.Pathology of Tumours of the Nervous System, 5th edition, Baltimore: Williams and Wilkins, pp. 809–54.

    Google Scholar 

  8. Coleman RE and Purohit OR, 1993, Osteoclast inhibition for the treatment of bone metastases.Cancer Treat Rev,19, 79–103.

    Google Scholar 

  9. McCutcheon IE, Baranco RA, Katz DA and Saris SC, 1990, Adoptive immunotherapy of intracerebral metastases in mice.J Neurosurgery,72, 102–9.

    Google Scholar 

  10. Budd GT, Green S, Baker LH, Hersh EP, Weick JK and Osborne CK, 1991, A Southwest Oncology Group phase II trial of recombinant tumor necrosis factor in metastatic breast cancer.Cancer,68, 1694–5.

    PubMed  Google Scholar 

  11. Fidler IJ and Schackert G, 1991, Blood-brain barrier and pathogenesis of experimental brain metastasis.Cancer Bull,43, 33–40.

    Google Scholar 

  12. Olden K, 1990, Human tumor bone metastasis model in athymic nude rats.JNCI,82, 340–1.

    PubMed  Google Scholar 

  13. Price JE and Zhang RD, 1990, Studies of human breast cancer metastasis using nude mice.Cancer Metastasis Rev,8, 285–97.

    PubMed  Google Scholar 

  14. Nicolson GL and Poste G, 1983, Tumor implantation and invasion at metastatic sites.Int Rev Exp Pathol,25, 77–181.

    PubMed  Google Scholar 

  15. Rudland PS, Dunnington DJ, Kim U,et al. 1989, Isolation and properties of cell lines from the metastasizing rat mammary tumor SMT-2A.Br J Cancer,59, 854–64.

    PubMed  Google Scholar 

  16. Kjonniksen I, Hoifodt HK and Fodstad O, 1991, Different metastatic patterns of a human melanoma cell line in nude mice and rats: influence of microenvironment.JNCI,83, 1020–24.

    PubMed  Google Scholar 

  17. Nicolson GL, 1984, Cell surface molecules and tumor metastasis. Regulation of metastatic phenotypic diversity.Exp Cell Res,150, 3–22.

    PubMed  Google Scholar 

  18. Miller FR, 1993, Immune mechanisms in the sequential steps of metastasis.Crit Rev Oncogenesis,4, 293–311.

    PubMed  Google Scholar 

  19. Markovic SN and Murasko DM, 1991, Role of natural killer and T-cells in interferon-induced inhibition of spontaneous metastasis of the B16F10L murine melanoma.Cancer Res,51, 1124–8.

    PubMed  Google Scholar 

  20. Sala A, Gresser I, Chassoux D,et al. 1992, Inhibition of Friend leukemia cell visceral metastases by a new monoclonal antibody and the role of the immune system of the host in its action.Cancer Res,52, 2880–9.

    PubMed  Google Scholar 

  21. Jung A, Bornand J, Mermillod B, Edouard C and Meunier PJ, 1984, Inhibition by diphosphonates of bone resorption induced by the Walker tumor of the rat.Cancer Res,44, 3007–11.

    PubMed  Google Scholar 

  22. Arguello F, Baggs RB, Duerst RE,et al. 1990, Pathogenesis of vertebral metastasis and epidural spinal cord compression.Cancer,65, 98–106.

    PubMed  Google Scholar 

  23. Inoue T, Steck PA and Nicolson GL, 1990, Increased expression of transferrin-like receptors and response to transferrin are related to brain-metastasizing properties of rat mammary adenocarcioma sublines.Proc Am Assoc Cancer Res,41, 54 (Abstract).

    Google Scholar 

  24. Manzotti C, Audisio RA and Pratesi G, 1993, Importance of orthotopic implantation for human tumors as model systems: relevance to metastasis and invasion.Clin Exp Metastasis,11, 5–14.

    PubMed  Google Scholar 

  25. Arguello F, Baggs RB, Eskenazi AE, Duerst RE and Frantz CN, 1991, Vascular anatomy and organ-specific tumor growth as critical factors in the development of metastases and their distribution among organs.Int Cancer,48, 1–8.

    Google Scholar 

  26. Shevrin DH, Kukreja C, Ghosh L and Lad TE, 1988, Development of skeletal metastasis by human prostate cancer in athymic nude mice.Clin Exp Metastasis,6, 401–9.

    PubMed  Google Scholar 

  27. Kjonniksen I, Nesland JM, Pihl A and Fodstad O, 1990, Nude rat model for studying metastasis to bone and bone marrow.JNCI,82, 408–12.

    PubMed  Google Scholar 

  28. Arguello F, Frantz CN and Baggs RB, 1990, Experimental bone and bone marrow metastasis in laboratory animals (letter).JNCI,82, 1069–70.

    PubMed  Google Scholar 

  29. Russo J and Russo IH, 1991, Mammary tumorigenesis. In: Ito N and Sugano H, eds.Modification of Tumor Development in Rodents. Progress in Experimental Tumor Research, volume 33. New York: Karger, pp. 175–91.

    Google Scholar 

  30. Stoica G, Koestner A and Capen CC, 1983, Characterization of N-ethyl-N-nitrosourea-induced mammary tumors in the rat.Am J Pathol,110, 161–9.

    PubMed  Google Scholar 

  31. Druckrey H, 1971, Genotypes and phenotypes of ten inbred strains of BD-rats.Arzneim-Forsch (Drug Res),8, 1274–8.

    Google Scholar 

  32. PPatterson MK, 1979, Measurement of growth and viability of cells in culture.Methods Enzymol,58 141–52.

    PubMed  Google Scholar 

  33. Sheehan DC and Hrapchak BB, eds,Theory and Practice of Histotechnology, second edition, pp. 59–159. St Louis: C. V. Mosby Co, 1980.

    Google Scholar 

  34. Karnovsky MJ, 1965, A formaldehyde-glutaraldehyde fixative of high osmolality for use in electron microscopy.J Cell Biol,27, 137A-138A.

    Google Scholar 

  35. Hayat MA, ed,Principles and Techniques of Electron Microscopy, Biological Applications, third edition. Boca Raton: CRC Press, Inc. 1989.

    Google Scholar 

  36. Tate EH, Wilder ME, Cram LS and Wharton W, 1983, A method for staining 3T3 cell nuclei with propridium iodide in hypotonic solution.Cytometry,4, 211–15.

    PubMed  Google Scholar 

  37. Hiddemann W, Schumann J, Andreeff M,et al. 1984, Convention on nomenclature for DNA cytometry.Cytometry,5, 445–6.

    Google Scholar 

  38. Ott L.An Introduction to Statistical Methods and Data Analysis, third edition. pp. 124–216. Boston: PWS-Kent Publishing Co., 1988.

    Google Scholar 

  39. Stoica G, Koestner A and O'Leary M, 1985, Characteristics of normal rat mammary epithelial cells and N-ethyl-N-nitrosourea-induced mammary adenocarcinoma cells grown in culture.Anticancer Res,5, 499–510.

    PubMed  Google Scholar 

  40. Dressler LG, Seamer LC, Owens MA, Clark GM and McGuire WL, 1988, DNA flow cytometry and prognostic factors in 1331 frozen breast cancer specimens.Cancer,61, 420–7.

    PubMed  Google Scholar 

  41. Fidler IJ and Kripke ML, 1977, Metastasis results from preexisting variant cells within a malignant tumor.Science,197, 893–5.

    PubMed  Google Scholar 

  42. Raz A and Hart IR, 1980, Murine melanoma: a model for intracranial metastasisBritish J Cancer,42, 331–41.

    Google Scholar 

  43. Tao T, Matter A, Vogel K and Burger M, 1979, Liver-colonizing melanoma cells selected from B-16 melanoma.Int J Cancer,23, 854–7.

    PubMed  Google Scholar 

  44. Brunson KW and Nicolson GL, 1979, Selection of malignant melanoma variant cell lines for ovary colonization.J Supramol Struct,11, 517–28.

    PubMed  Google Scholar 

  45. Miner K, Kawaguchi T, Uba GW and Nicolson GL, 1982, Clonal drift of cell surface, melanogenic, and experimental metastatic properties ofin vivo-selected, brain meninges-colonizing murine B16 melanoma.Cancer Res,42, 4631–6.

    PubMed  Google Scholar 

  46. Stackpole CW, Alterman AL and Valle EF, 1991, B16 melanoma variants selected by one or more cycles of spontaneous metastasis to the same organ fail to exhibit organ specificity.Clin Exp Metastasis,9, 319–32.

    PubMed  Google Scholar 

  47. Shearman P and Longenecker B, 1980, Selection for virulence and organ-specific metastasis of herpes-virus-transformed lymphoma cells.Int J Cancer,25, 363–9.

    PubMed  Google Scholar 

  48. Neri A, Welch D, Kawaguchi T and Nicolson GL, 1982, Development and biologic properties of malignant cell sublines and clones of a spontaneously metastasizing rat mammary adenocarcinoma.JNCI,68, 507–17.

    PubMed  Google Scholar 

  49. Nicolson GL, 1988, Differential organ tissue adhesion, invasion, and growth properties of metastasizing rat mammary adenocarcinoma cells.Breast Cancer Res Treat,12, 167–76.

    PubMed  Google Scholar 

  50. Zetter BR, 1990, The cellular basis of site-specific metastasis.New Eng J Med,322, 605–12.

    PubMed  Google Scholar 

  51. Ozzello L. Benign and malignant proliferations of the mammary epithelium: ultrastructural observations. In: McDivitt RW, Oberman HA, Ozello L and Kaudman N, eds.The Breast. pp. 106–38, Baltimore: Williams and Wilkins, 1984.

    Google Scholar 

  52. Dolezalova V, Nemacek R, Toth J and Sugar J, 1987, Phenotypic alterations of 7,12-dimethylbenz-(a)anthracene and N-methy-N-nitrosourea induced rat mammay tumors during repeated transplantations.In Vivo,1, 257–64.

    PubMed  Google Scholar 

  53. Bae SN, Arand G, Azzam H,et al. 1993, Molecular and cellular analysis of basement membrane invasion by human breast cancer cells in matrigel-basedin vitro assays.Breast Cancer Res Treat,24, 241–55.

    PubMed  Google Scholar 

  54. Frankfurt OS, Slocum HK, Rustum YM,et al. 1984, Flow cytometric analysis of DNA aneuploidy in primary and metastatic human solid tumors.Cytometry,5, 71–80.

    PubMed  Google Scholar 

  55. Gijbels MJJ, Visser JWM, Solleveld HA, Broerse JJ and Zurcher C, 1991, Flow cytometric DNA measurement and cytomorphometric analysis of formalin-fixed rat mammary tumors.Brit J Cancer,64, 523–7.

    PubMed  Google Scholar 

  56. Parsons DF, 1989, Near-diploid tumor cell lines for cancer research.Cancer Cells,1, 96–8.

    PubMed  Google Scholar 

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Authors and Affiliations

  1. Department of Veterinary Pathology, The University of Georgia, Athens, GA

    D. Greg Hall

  2. Department of Veterinary Pathobiology, Texas A & M University, TX, USA

    G. Stoica

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Hall, D.G., Stoica, G. Characterization of brain and bone-metastasizing clones selected from an ethylnitrosourea-induced rat mammary carcinoma. Clin Exp Metast 12, 283–295 (1994). https://doi.org/10.1007/BF01753835

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