Skip to main content
Log in

Comparative morphology and tumourigenicity of human hepatocellular carcinoma cell lines in athymic rats and mice

  • Published:
Virchows Archiv A Aims and scope Submit manuscript

Summary

Four human hepatoma cell lines PLC/PRF/5, Hep G2, Sk-Hep 1 and Mahlavu were inoculated subcutaneously into athymic Balb/c nude mice and N/NIH outbred nude rats, producing well encapsulated tumours. The 4 hepatoma tumour types in the athymic rodents differ morphologically. PLC/PRF/5 and Hep G2 cells are well differentiated polygonal cells which resemble normal hepatocytes. Tumour arrangement is characterized by solid masses and trabeculae while stromal support is minimal. In contrast, Mahlavu and Sk-Hep 1 tumours have a sarcomatous appearance and consist of spindle shaped cells arranged in solid masses with a rich stromal support. Tumourigenicity of hepatoma cells in the athymic rodents was dependent on injected cell type, inoculation density, relative immunocompetence of the host and the species of animals used. In nude mice, Sk-Hep 1 cells were the most tumourigenic, while Hep G2 cells were tumourigenic only at very high inoculation densities. In nude rats, which were more resistant to tumour formation, PLC/PRF/5 cells were the most tumourigenic. Pre-treatment of athymic mice and rats with total body irradiation resulted in enhanced tumourigenicity for all hepatoma cell lines tested. This was manifested as increased “take” rates, a decreased latency from tumour cell injection to tumour detection, increased tumour weight, and for PLC/PRF/5 cells an increased invasiveness to adjacent body cavities. Furthermore, following irradiation, the minimal number of injected cells required to produce subcutaneous tumours was markedly reduced in both animal species, regardless of tumour cell type. The protocols described enable the reproduceable growth of human hepatoma tumours in athymic rodents.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Similar content being viewed by others

References

  • Aden DP, Fogel A, Plotkin S, Damjanov I, Knowels BB (1979) Controlled synthesis of HBsAg in a differentiated human liver carcinoma cell line. Nature 282:615–616

    Google Scholar 

  • Alexander JJ, MacNab G, Saunders R (1978) Studies on in vitro production of hepatitis B surface antigen by a human hepatoma cell line. Perspec Virol 10:103–117

    Google Scholar 

  • Bassendine MF, Arborgh BAM, Shipton N, Monjardino J, Aranguibel F, Thomas HC, Sherlock S (1980) HBsAg and alpha feto protein secreting human primary liver cell cancer in athymic mice. Gastroenterology 79:528–532

    Google Scholar 

  • Brooks CG, Webb PJ, Robins RA, Robinson G, Baldwin RW, Festing MFW (1980) Studies on the immunobiology of rnu/rnu nude rats with congenital aplasia of the thymus. Eur J Immunol 10:58–65

    Google Scholar 

  • Carlson RI, Ben Porath E, Shouval D, Strauss W, Isselbacher KJ, Wands JR (1985) Antigenic characterization of human hepatocellular carcinoma. Development of in vitro and in vivo immunoassays that use monoclonal antibodies. J Clin Invest 76:40–51

    Google Scholar 

  • Chang R (1954) Continuous subcultivation of epithelial like cells from normal human tissue. Proc Soc Exp Biol NY 87:440–443

    Google Scholar 

  • Chen JN (1964) Establishment in vitro and some preliminary observations on a strain of human liver cell carcinoma. Acta Unio Int Contra Carcinoma 20:1314–1315

    Google Scholar 

  • Chisari FV, Bieber MS, Johnson CA, Xavier C, Anderson DA (1981) Functional properties of lymphocyte subpopulations in hepatitis B virus infection. J Immunol 126:45–49

    Google Scholar 

  • Colston MJ, Fieldsteel AH, Dawson PJ (1981) Growth and regression of human tumour cell lines in congenitally athymic (rnu/rnu) rats. JNCO 66:843–847

    Google Scholar 

  • Colston MJ, Fieldsteel AH, Lancaster RD, Dawson PJ (1982) Immunologic studies and growth of human tumours in the athymic rat. In: Proceedings of the 3rd International Workshop on Nude Mice, Fischer, New York, pp 189–196

    Google Scholar 

  • Das PK, Nayak NC, Tsiquaye KN, Zuckerman AJ (1980) Establishment of a human hepatocellular carcinoma cell line releasing hepatitis B virus surface antigen. Br J Exp Pathol 61:648–654

    Google Scholar 

  • Doi I, Namba M, Sato J (1975) Establishment and some biologic characteristics of human hepatoma cell lines. Gann 66:385–392

    Google Scholar 

  • Festing MFW, May D, Connors TA, Lowell D, Sparrow S (1978) An athymic nude mutation in the rat. Nature 274:365–366

    Google Scholar 

  • Fogh J, Wright WC, Loveless JD (1977a) Absence of HeLa cell contamination in 169 cell lines derived from human tumours. J Natl Cancer Inst 58:209–214

    Google Scholar 

  • Fogh J, Fogh JM, Orefeo T (1977b) One hundred and twenty seven cultured human tumour cell lines producing tumours in nude mide. J Natl Cancer Inst 59:221–226

    Google Scholar 

  • Giovanella BC, Stehlin JS, Williams LJ Jr, Lee SS, Shepard RC (1978) Heterotransplantation of human cancers into nude mice. Cancer 42:2269–2281

    Google Scholar 

  • Habu S, Fukui H, Shinamura K, Kasai M, Nagai Y, Okumura K, Tamoki N (1981) In vivo effects of anti-asialo GM1. I. Reduction of NK activity and enhancement of transplanted tumour growth in nude mice. J Immunol 127:34–38

    Google Scholar 

  • Hirohashi S, Shimosato Y, Kameya T, Koide T, Mukojima T, Taguchi Y (1977) Morphological and functional aspects of human liver cell carinomas transplanted in nude mice. Proc 2nd Int Workshop on nude mice, University of Tokyo Press, Tokyo, pp 427–434; G Fischer, Stuttgart

    Google Scholar 

  • Hirohashi S, Shimosato Y, Kameya T, Koide T, Mukojima T, Taguchi Y, Keizo Kageyama (1979) Production of alpha feto protein and normal serum proteins by xenotransplanted human hepatomas in relation to their growth and morphology. Cancer Res 39:1819–1828

    Google Scholar 

  • Kintzen W, Silny J (1960) Peliosis hepatis after administration of fluoxymesterone. Can Med Assoc J 83:860–862

    Google Scholar 

  • Knowles BB, Adem DP (1980) Human hepatocellular carcinoma cell lines secrete the major plasma proteins and hepatitis B surface antigen. Science (USA) 209:497–499

    Google Scholar 

  • Lapis K (1983) Morphologic expression of neoplasia in human and experimental liver tumours. In: 13th Int Cancer Congress, Part D, Research and Treatment, Riss, New York, pp 67–76

    Google Scholar 

  • Lotzova E, Savary CA, Gray KN, Raulston GL, Jardine JH (1984) NK cell profile of two random bred strains of athymic rats. Exp Hematol 12:633–640

    Google Scholar 

  • Macnab GM, Alexander JJ, Lecatsas G, Bey EM, Urabanowich JM (1976) Hepatitis B surface antigen produced by a human hepatoma cell line. Br J Cancer 34:509–515

    Google Scholar 

  • McDowell EM (1978) Fixation and Processing. In: Trump BF, Jones RT (eds) Diagnostic electron microscopy. Whey and Sons, New York, 1:pp 113–119

    Google Scholar 

  • Nakabayashi H, Takketa K, Yamane T, Miyazeke M, Miyano K, Sato J (1984) Phenotypical state of a human hepatoma cell line HuH-7 in long term culture with chemically defined medium. Gann 75:151–158

    Google Scholar 

  • Owens R, Smith H, Nelson-Rees W, Springer EL (1976) Brief communication: Epithelial cell cultures from normal and cancerous human tissues. J Natl Cancer Inst 56:843–846

    Google Scholar 

  • Prozesky OW, Brits CJ, Grabow WOK (1972) In vitro culture of cell lines from Australia antigen positive and negative patients. In: Saunders SJ, Terblanche J (eds) Liver, Pitman Medical Publications, London, pp 358–360

    Google Scholar 

  • Reid LM, Minato N, Gresser I, Holland J, Kadish A, Bloom R (1981) Influence of anti-mouse interferon serum on the growth and metastasis of tumour cells persistently infected with virus and of human prostatic tumours in athymic nude mice. Proc Nat Acad Sci USA 78:1171–1175

    Google Scholar 

  • Sattler FR, Paolucci S, Kreider JW, Ladda RL (1982) A human hepatoma cell line (PLC/PRF/5) produces lung metastases and secretes HBsAg in nude mice. Eur J Cancer Clin Oncol 18:381–389

    Google Scholar 

  • Shouval D, Reid LM, Chakraborty PR, Ruiz-Opazo N, Morecki R, Gerber MA, Thung SN, Shafritz DA (1981) Tumourigenicity in nude mice of a human hepatoma cell line containing hepatitis B virus DNA. Cancer Res 41:1342–1350

    Google Scholar 

  • Shouval D, Shafritz DA, Zurawski VR Jr, Wands JR (1982a) Immunotherapy in nude mice of a human hepatoma using monoclonal antibodies against hepatitis B virus. Nature (Lond) 298:567–569

    Google Scholar 

  • Shouval D, Wands JR, Zurawski VR Jr, Isselbacher KJ, Shafritz DA (1982b) Protection against experimental hepatoma formation in nude mice by monoclonal antibodies to hepatitis B virus antigen. Hepatology 2:128s-133s

    Google Scholar 

  • Shouval D, Rager-Zisman B, Quan P, Shafritz DA, Bloom BR, Reid LM (1983) Role in nude mice of interferon and NK cells in inhibiting the tumourigenicity of human hepatocellular carcinoma cells infected with heaptitis B virus. J Clin Invest 72:707–717

    Google Scholar 

  • Shouval D, Eilat D, Carlson RI, Adler R, Livni N, Wands JR (1985) Human hepatoma associated cell surface antigen: Identification and characterization by means of monoclonal antibodies. Hepatology 5:347–356

    Google Scholar 

  • Thomas HC, Montano L, Goodall A, de Koning R, Oledapo J, Wiedman KH (1982) Immunological mechanisms in chronic hepatitis B virus infection. Hepatology 2:116s-121s

    Google Scholar 

  • Watanabi T, Morizane T, Tsuchimotok K, Inagaki Y, Munakata Y, Nakamura T, Kumagai N, Tsuchiya M (1983) Establishment of a cell line (HCC-M) from a human hepatocellular carcinoma. Int J Cancer 32:141–146

    Google Scholar 

  • Zak FG (1950) Peliosis hepatis. Am J Pathol 26:1–15

    Google Scholar 

Download references

Author information

Authors and Affiliations

Authors

Rights and permissions

Reprints and permissions

About this article

Cite this article

Shouval, D., Schuger, L., Levij, I.S. et al. Comparative morphology and tumourigenicity of human hepatocellular carcinoma cell lines in athymic rats and mice. Vichows Archiv A Pathol Anat 412, 595–606 (1988). https://doi.org/10.1007/BF00844296

Download citation

  • Accepted:

  • Issue Date:

  • DOI: https://doi.org/10.1007/BF00844296

Key words

Navigation