Abstract
Assessment of the function of putative dominantly-acting oncogenes or recessive tumor-suppressor genes in human tumor development and progression must ultimately involve xenografting experiments using immune deficient animals such as nude mice. Most human tumor xenograft experiments have employed conventional subcutaneous injection procedures. However, despite the simplicity of this procedure, it poses some serious potential drawbacks as most types of human tumor will not readily grow or metastasize from a subcutaneous (‘ectopic’) site of injection. In contrast, ‘orthotopic’ injection procedures will often enhance the tumorigenic and/or metastatic ability of tumor cell populations. An example of this is summarized in the context of human malignant melanoma where the effects of subcutaneous versus subdermal injection are compared. Despite the seeming subtle and minor change in injection site, superior growth of human melanomas can be obtained by the latter, orthotopic-like, route of injection.
It therefore follows that induction of tumorigenic or metastatic properties in a given human cell population by gene transfection may not be detected if the transfected cells are assayed in vivo only by subcutaneous injection procedures. An example of this is provided by experiments involving transfection of normal or mutated ras genes into a low-grade, well-differentiated human bladder carcinoma cell line, called RT-4. Thus overexpression of normal or mutated (valine 12) c-H-ras resulted in acquisition of a clinical-like invasive phenotype. However, this was clearly seen only if the cells were injected into the bladders (i.e. ‘intravesically’) of nude mice. In contrast, conventional subcutaneous injection of the high ras expressing transfected RT-4 cell lines did not reveal acquisition of invasive properties: all cell lines grew locally as well-encapsulated tumor masses.
It is argued that similar orthotopic injection procedures should be employed when assessing the suppressive effects of various wild-type tumor-suppressor genes on human tumor growth in vivo. Utilization of subcutaneous injection procedures may grossly exaggerate the growth suppressive effects of such genes. This could explain the paradox of why, on the one hand, alterations involving many different genes (including different suppressor genes) appear to be involved in human carcinoma tumorigenesis while on the other hand, complete suppression of tumorigenicity can be caused by transfer of a single wild-type suppressor gene. Such complete suppressions might be observed only after ectopic (usually subcutaneous) injection procedures.
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References
Fearon ER, Vogelstein B: A genetic model for colorectal carcinogenesis. Cell 61: 759–767, 1990
Bishop JM: The molecular genetics of cancer. Science 234: 305–311, 1987
Mikkelsen T, Cavanee WK: Suppressors of the malignant phenotype. Cell Growth & Differentiation 1: 201–207, 1990
Gunthert U, Hofmann M, Rudy W, Reber S, Zoller M, Haussmann I, Matsku S, Wenzel A, Ponta H, Herrlich P: A new variant of glycoprotein CD-44 confers metastatic potential to rat carcinoma cells. Cell 65: 13–24, 1991
Vleminckx K, Vakaet L, Marcel M, Fiers W, Van Roy F: Genetic manipulation of E-cadherin expression by epithelial tumor cells reveals an invasion suppressor role. Cell, in press, 1991
Cajot J-F, Schleuning W-D, Medcalf RL, Bamat J, Testuz J, Lieberman L, Sordat B: Mouse L cells expressing prourokinase-type plasminogen activator: effects on extracellular matrix degradation and invasion. J Cell Biol 109: 915–925, 1989
Raz A, Pazerini G, Carmi P: Identification of the metastasis-associated galactoside-binding lectin as a chimeric gene product with homology to an IgE-binding protein. Cancer Res 49: 3489–3493, 1989
Leone A, Flatow U, King CR, Sandeen MA, Margulies IMK, Liotta LA, Steeg PS: Reduced tumor incidence, metastatic potential, and cytokine responsiveness of nm 23-transfected melanoma cells. Cell 65: 25–35, 1991
Hart IR, Easty D: Identification of genes controlling metastatic behaviour. Br J Cancer 63: 9–12, 1991
Wu D-D, Huszar D, Dick JE, Bernstein A, Phillips RA: High efficiency gene transfer and expression in normal murine B lymphocytes. J Immunol 101: 279–285, 1987
Sharkey FE, Fogh J: Considerations in the use of nude mice for cancer research. Cancer Metastasis Reviews 3: 341–360, 1984
Takahashi T, Nau MM, Chiba I, Birrer MJ, Rosenberg RK, Vincour M, Levitt M, Pass H, Gazdar AF, Minna JD: p 53: a frequent target for genetic abnormalities in lung cancer. Science 246: 491–494, 1988
Fidler IJ: Rationale and methods for the use of nude mice to study the biology and therapy of human cancer metastasis. Cancer Metastasis Reviews 5: 29–491, 1986
Fidler IJ, Naito S, Pathak S: Orthotopic implantation in essential for the selection, growth and metastasis of human renal cell cancer in nude mice. Cancer Metastasis Reviews 9: 145–165, 1990
Fidler IJ: Critical factors in the biology of human cancer metastasis: Twenty-eighth G.H.A. Clowes Memorial Award lecture. Cancer Res 50: 67130–6138, 1990
Ibrahiem EHI, Nigam VN, Brailovsky CA, Madarnas P, Elhilali M: Orthotopic implantation of primary N-[-4-Nitro-2 furyl)-2-thiazolyl} formanide-induced bladder cancer in bladder submucosa: an animal model for bladder cancer study. Cancer Res 43: 617–622, 1983
Miller FR, Medina D, Heppner GH: Preferential growth of mammary tumors in intact mammary fatpads. Cancer Res 41: 3863–3867, 1981
Bresalier RS, Hujanen ES, Raper SE, Roll FJ, Hzkowitz SH, Martin GR and Kim YS: An animal model for colon cancer metastasis: establishment and characterization of murine cell lines with enhanced liver-metastasizing ability. Cancer Res 47: 1398–1406, 1987
Naito S, von Eschensach AC, Giavazzi R, Fidler IJ: Growth and metastasis of tumor cells isolated from a human renal cell carcinoma implanted into different organs of nude mice. Cancer Res 46: 4109–4115, 1986
Gallie BL, Albert DM, Wong JJY, Buyukmichi N, Puliafito CA: Heterotransplantation of retinoblastoma into the athymic ‘nude’ mouse. Invest Ophthalmol 16: 256–259, 1977
Gallie BL, Chew EY, Chang M, Phillips RA: Retinoblastoma in the eyes of nude mice: quantitative assessment of therapy. In: Proc 3rd Int Workshop in Nude Mice, Gustav Fischer, New York 1982, pp 641–647
McLemore TL, Liu MD, Blacker PC, Gregg M, Alley MC, Abbott BJ, Shoemaker RH, Bohlman ME, Litterst CC, Hubard WC, Brennan RH, McMahon JB, Fine DL, Eggleston JC, Mayo JG, Boyd MR: Novel intrapulmonary model for orthotopic propogation of human lung cancers in athymic nude mice. Cancer Res 47: 5132–5140, 1987
Morikawa K, Walker SM, Nakajima M, Pathak S, Jessup JM, Fidler IJ: Influence of organ environment on the growth, selection, and metastasis of human colon carcinoma cells in nude mice. Cancer Res 48: 6863–6871, 1988
Morikawa K, Walker SM, Jessup JM, Fidler IJ: In vivo selection of highly metastatic cells from surgical specimens of different primary colon carcinomas implanted into nude mice. Cancer Res 48: 1943–1948, 1988
Jessup JM, Giavazzi R, Campbell D, Cleary KR, Morikawa K, Hostetter R, Atkinson EN, Fidler IJ: Metastatic potential of human colorectal carcinomas implanted into nude mice: prediction of clinical outcome in patient operated for cure. Cancer Res 49: 6906–6910, 1989
Cornil I, Man MS, Fernandez B, Kerbel RS: Enhanced tumorigenicity, melanogenesis and metastasis of a human malignant melanoma observed after subdermal implantation in nude mice. J Natl Cancer Inst 81: 938–944, 1989
Theodorescu D, Cornil I, Fernandez BJ, Kerbel RS: Over-expression of normal and mutated forms of HRAS induces orthotopic bladder invasion in a human transitional cell carcinoma. Proc Natl Acad Sci USA 87: 9047–9051, 1990
Giovenella BC, Yim SO, Stehlin JS: Development of invasive tumors in the ‘nude’ mouse after injection of cultured human melanoma cells. J Natl Cancer Inst 48: 1531–1533, 1972
Giovenella BC, Yim SO, Morgan AC, Stehlin JS, Williams LJ: Metastases of human melanomas transplanted in ‘nude’ mice. J Natl Cancer Inst 50: 1051–1053, 1973
Pawlowski A, Lea PJ: Human melanoma xenografts in skin tumors: In: Experimental and Clinical Aspects Cont CJ, et al (eds), Raven Press, New York, 1989, pp 103–132
Sordat BCM, Ueyama Y, Fogh J: Metastasis of tumor xenografts in the nude mouse. In: The Nude Mouse in Experimental and Clinical Research, Fogh J and Giovenella BC (eds) New York Academic Press, 1982, Vol 2 pp 95–147
Kerbel RS, Man MS, Dexter D: A model of human cancer metastasis: extensive spontaneous and artificial metastasis of a human pigmented melanoma and derived variant sublines in nude mice. J Natl Cancer Inst 72: 93–108, 1984
Kozlowski JM, Hart IR, Fidler IJ, Hauna NA. A human melanoma line heterogeneous with respect to metastatic capacity in athymic nude mice. J Natl Cancer Inst 72: 913–917, 1984
Fodstad O, Kjonniksen I, Aandal S. Nesland JM, Boyd MR, Pilh A: Extrapulmonary tissue-specific metastasis formation in nude mice injected with FEMX-I human melanoma cells. Cancer Res 48: 4382–4388, 1988
Welch ER, Bisi JE, Miller BE, Conaway D, Seftor EA, Yohem KH, Gilmore LM, Seftor REB, Nakajima M, Hendrix M: Characterization of highly invasive and spontaneously metastatic human malignant melanoma cell line. Int J Cancer 47: 227–237, 1991
Mueller BM, Romerdahl CA, Trent JM, Reisfeld RA: Suppression of spontaneous melanoma metastasis in SCID mice with an antibody to the epidermal growth factor receptor. Cancer Res in press, 1991
Lockshin A, Giovenella BC, Delpoly PD, Williams LJ, Mendoza JT, Yim SO, Stehlin JS: Exceptional lethality for nude mice of cells derived from a primary human melanoma. Cancer Res 45: 345–350
Amer MH, Al-Sarraf M, Baker LH, Vaitkevicius M: Malignant melanoma and central nervous system metastases: incidence, diagnosis, treatment and survival. Cancer 42: 660–668, 1978
Ishikawa M, Fernandez B, Kerbel RS: Highly pigmented human melanoma variant which metastasizes widely in nude mice, including to skin and brain. Cancer Res 48: 4897–4903, 1988
Ishikawa M, Dennis JW, Man MS, Kerbel RS: Isolation and characterization of spontaneous wheat germ agglutinin-resistant (WGAr) human melanoma mutants displaying remarkably different metastatic profiles in nude mice. Cancer Res 48: 665–670, 1988
Ishikawa M, Kerbel RS: Characterization of a metastasis-deficient lectin resistant human melanoma mutant. Int J Cancer 43: 134–139, 1989
Clark WH, Elder DE, Van Horn M: The biologic forms of malignant melanoma. Human Path 17: 443–450, 1986
Herlyn M: Human melanoma: development and progression. Cancer Metastasis Reviews 9: 102–112, 1990
Kerbel RS: The growth dominant metastatic phenotype: cellular and molecular aspects. Adv Cancer Res 55: 87–132, 1990
Kath R, Rodeck U, Parmiter A, Jambrosic J, Herlyn M: Growth factor independence in vitro of primary melanoma cells from advanced but not early or intermediate lesions. Cancer Therapy Control 1: 179–191, 1990
Cornil I, Theodorescu D, Man MS, Herlyn M, Jambrosic J, Kerbel RS: Fibroblast cell interactions with human melanoma cells affect tumor cell growth as a function of tumor progression. Proc Natl Acad Sci (USA) 88: 6028–6032, 1991
Kerbel RS, Waghorne C, Korczak B, Lagarde A, Breitman M: Clonal dominance of primary tumors by metastatic tumor cells: genetic analysis and biological implications. Cancer Surveys 7: 597–629, 1988
Sidransky D, Von Eschenbach A, Tsai YC, Jones P, Summerhayes I, Marshall F, Paul M, Green P, Hamilton SR, Frost P, Vogelstein B: Identification of p 53 gene mutations in bladder cancers and urine samples. Science 252: 706–709, 1991
Smith K, Fennelly JA, Neal DE, Hall RR, Harris RR, Harris AL: Characterization and quantitation of the epidermal growth factor receptor in invasive and superficial bladder tumors. Cancer Res 49: 5810–5815, 1989
Messing EM: Clinical implications of the expression of epidermal growth factor receptors in human transitional cell carcinoma. Cancer Res 50: 2530–2537, 1990
Neal DE, Sharples L, Smith K, Fennelly J, Hall RR, Harris AL: The epidermal growth factor receptor and the prognosis of bladder cancer. Cancer 65: 1619–1625, 1990
Ahrling TE, Dubeau L, Jones PA: A new in vitro model to study invasion and metastasis of human bladder carcinomas. Cancer Res 47: 6660–6665, 1987
Saxon PJ, Srivatsan ES, Stanbridge E: Introduction of human chromosome 11 via microcell transfer controls tumorigenic expression of HeLa cells. EMBO J 5: 3461–3466, 1986
Tanaka T, Oshimura M, Kikuchi R, Seki M, Hayashi T, Miyaki M: Suppression of tumorigenicity in human colon carcinoma cells by introduction of normal chromosome 5 or 18. Nature 349: 340–342, 1991
Huang H-JS, Yees J-K, Shew J-Y, Chen P-L, Bookstein R, Freidmann T, Lee EY-HP, Lee WM: Suppression of the neoplastic phenotype by replacement of the Rb gene in human cancer cells. Science 242: 1563–1566, 1988
Bookstein R, Shew J-Y, Chen PL, Scully P, Lee WM: Suppression of tumorigenicity of human prostate carcinoma cells by replacing a mutated RB gene. Science 247: 712–715, 1990
Trent JM, Stanbridge E, McBride HL, Meese E, Casey G, Araujo DE, Witkowski CM, Nagel RB: Tumorigenicity in human melanoma cells controlled by introduction of human chromosome 6. Science 247: 568–571, 1986
Weissman BE, Saxon PJ, Pasquale SR, Jones GR, Geiser A, Stanbridge EJ: Introduction of normal human chromosome 11 into a Wilms tumor cell line controls its tumorigenic conversion. Science 236: 252–259, 1987
Fridman R, Giaccione G, Kanemoto T, Martin GR, Gazdar AF, Mulshine JL: Reconstituted basement membrane (matrigel) and laminin can enhance the tumorigenicity and the drug persistence of small cell lung cancer cell lines. Proc Natl Acad Sci USA 86: 6698–6702, 1990
Mule JJ, Jicha DL, Aebersold PM, Travis WD, Rosenberg SA: Disseminated human malignant melanoma in congenitally immune-deficient (bg/m/xid) mice. J Natl Cancer Inst 83: 350–355, 1991
Camps JL, Chang SM, Hsu TC, Freeman MR, Hong SJ, Zhau HE, von Eschenbach A, Chung LWK: Fibroblast-mediated acceleration of human epithelial tumor growth in vivo. Proc Natl Acad Sci USA 87: 75–79, 1990
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Departments of Medical Biophysics & Medical and Molecular Genetics, University of Toronto
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Kerbel, R.S., Cornil, I. & Theodorescu, D. Importance of orthotopic transplantation procedures in assessing the effects of transfected genes on human tumor growth and metastasis. Cancer Metast Rev 10, 201–215 (1991). https://doi.org/10.1007/BF00050792
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DOI: https://doi.org/10.1007/BF00050792