Advertisement

Virchows Archiv

, Volume 430, Issue 1, pp 29–35 | Cite as

Induction of apoptosis in a human hepatocellular carcinoma cell line by a neutralizing antibody to transforming growth factor-α

  • Shuichi Seki
  • Yasuhiko Sakai
  • Takuya Kitada
  • Nobuyoshi Kawakita
  • Atsushi Yanai
  • Hiroko Tsutsui
  • Hiroki Sakaguchi
  • Tetsuo Kuroki
  • Takeyuki Monna
Original Article

Abstract

A cell line derived from a Japanese man with hepatocellular carcinoma was established in culture and designated OCUH-16. The cell line has the morphological and chromosomal features of hepatocellular carcinoma cells and has a short doubling time (≈33 h). OCUH-16 cells were shown to express transforming growth factor-α, (TGF-α) in addition to albumin, DNA polymerase-α, c-JUN, and the retinoblastoma gene product. Electron microscopy revealed TGF-α immunoreactivity associated with the cell membrane, but TGF-α was not detected in medium conditioned by OCUH-16 cells by enzyme-linked immunosorbent assay. Reverse transcription and polymerase chain reaction analysis revealed the presence of TGF-α messenger RNA in these cells. Culture of OCUH-16 cells in the presence of a neutralizing antibody to TGF-α inhibited cell proliferation and induced many cells to undergo apoptosis (programmed cell death). These observations suggest that endogenous TGF-α is necessary for OCUH-16 cell growth.

Key words

Hepatocellular carcinoma Apoptosis Transforming growth factor-α 

Preview

Unable to display preview. Download preview PDF.

Unable to display preview. Download preview PDF.

References

  1. 1.
    Alcorn JA, Feitelberg SP, Brenner DA (1990) Transient induction of c-jun during hepatic regeneration. Hepatology 11:909–915Google Scholar
  2. 2.
    Anklesaria P, Teixido J, Laiho M, Pierce JH, Greenberger JS, Massague J (1990) Cell-cell adhesion mediated by binding of membrane-anchored transforming growth factor a to epidermal growth factor receptors promotes cell proliferation. Proc Natl Acad Sci USA 87:3289–3293Google Scholar
  3. 3.
    Barrandon Y, Green H (1987) Cell migration is essential for sustained growth of keratino-cyte colonies: the roles of transforming growth factor-α and epidermal growth factor. Cell 50:1131–1137Google Scholar
  4. 4.
    Bensch KG, Tanaka S, Hu S-Z, Wang TS-F, Korn D (1982) Intracellular localization of human DNA polymerase a with monoclonal antibodies. J Biol Chem 257:8391–8396Google Scholar
  5. 5.
    Brenner DA, Koch KS, Leffert HL (1989) Transforming growth factor-α stimulates proto-oncogene c-jun expression and a mitogenic program in primary cultures of adult rat hepatocytes. DNA 8:279–285Google Scholar
  6. 6.
    Bruix J, Barrera JM, Calvet X, Ercilla G, Costa J, Sanchez-Tapias JM, Ventura M, Vall M, Bruguera M, Bru C, Castillo R, Rodes J (1989) Prevalence of antibodies to hepatitis C virus in Spanish patients with hepatocellular carcinoma and hepatic cirrhosis. Lancet II:1004–1006Google Scholar
  7. 7.
    Castilla A, Prieto J, Fausto N (1991) Transforming growth factors β 1 and α in chronic liver disease: effects of interferon alpha therapy. N Engl J Med 324:933–940Google Scholar
  8. 8.
    Chomczynski P, Sacchi N (1987) Single-step method of RNA isolation by acid guanidinium thiocyanate-phenol-chloroform extraction. Anal Biochem 162:156–159Google Scholar
  9. 9.
    Choo Q-L, Kuo G, Weiner AJ, Overby LR, Bradley DW, Houghton M (1989) Isolation of a cDNA clone derived from a blood-borne non-A, non-B viral hepatitis genome. Science 244:359–362Google Scholar
  10. 10.
    Cohen JJ, Duke RC (1984) Glucocorticoid activation of a calcium-dependent endonuclease in thymocyte nuclei leads to cell death. J Immunol 132:38–42Google Scholar
  11. 11.
    Collins MKL, Lopes Rivas A (1993) The control of apoptosis in mammalian cells. Trends Biochem Sci 18:307–309Google Scholar
  12. 12.
    Colombo M, Kuo G, Choo QL, Donate ME, Del Ninno E, Tomasino MA, Dioguardi N, Houghton M (1989) Prevalence of antibodies to hepatitis C virus in Italian patients with hepatocellular carcinoma. Lancet 11:1006–1008Google Scholar
  13. 13.
    Dazza M-C, Meneses L-V, Girard P-M, Villaroel C, Brechot C, Larouze B (1990) Hepatitis C virus antibody and hepatocellular carcinoma. Lancet 335:1216Google Scholar
  14. 14.
    Derynck R (1992) The physiology of transforming growth factor-α. Adv Cancer Res 58:27–52Google Scholar
  15. 15.
    Dexter TM (1993) Synergistic interactions in haemopoiesis: biological implications and clinical use. Eur J Cancer [A] 29 [Suppl 3]:S6-S9Google Scholar
  16. 16.
    Edmondson HA, Steiner PE (1954) Primary carcinoma of the liver: a study of 100 cases among 48,900 necropsies. Cancer 7:462–503Google Scholar
  17. 17.
    Fukuda K, Kojiro M, Chin J-F (1993) Demonstration of extensive chromatin cleavage in transplanted Morris hepatoma 7777 tissue: apoptosis or necrosis? Am J Pathol 142:935–946Google Scholar
  18. 18.
    Hasan F, Jeffers LJ, De Medina M, Reddy KR, Parker T, Schiff ER, Houghton M, Choo QL, Kuo G (1990) Hepatitis CC associatedd hepatocellularr carcinoma. Hepatology 12:589–591Google Scholar
  19. 19.
    Hiramine C, Hojo K, Koseto M, Nakagawa T, Mukasa A (1990) Establishment of a murine thymic epithelial cell line capable of inducing both thymic nurse cell formation and thymocyte apoptosis. Lab Invest 62:41–54Google Scholar
  20. 20.
    Hsia CC, Axiotis CA, Di Bisceglie AM, Tabor E (1992) Transforming growth factor-alpha in human hepatocellular carcinoma and coexpression with hepatitis B surface antigen in adjacent liver. Cancer 70:1049–1056Google Scholar
  21. 21.
    Jhappan C, Stahle C, Harkins RN, Fausto N, Smith GH, Merlino TG (1990) TTGF-α overexpression in transgenic mice induces liver neoplasia and abnormal development of the mammary gland and pancreas. Cell 61:1137–1146Google Scholar
  22. 22.
    Kawano S, Fujiyama S, Sato S, Tanaka M, Goto M, Taura Y, Sato T, Kawahara T, Mizuno K, Nonaka S (1992) Clinical evaluation of three anti-HCV ELISA in patients with various liver diseases. Dig Dis Sci 37:1268–1274Google Scholar
  23. 23.
    Kerr JFR, Wyllie AH, Currie AR (1972) Apoptosis: a basic biological phenomenon with wide-ranging implications in tissue kinetics. Br J Cancer 26:239–257Google Scholar
  24. 24.
    Kuo G, Choo Q-L, Alter HJ, Gitnick GL, Redeker AG, Purcell RH, Miyamura T, Dienstag JL, Alter MJ, Stevens CE, Tegtmeier GE, Bonino F, Colombo M, Lee W-S, Kuo C, Berger K, Shuster JR, Overby LR, Bradley DW, Houghton M (1989) An assay for circulating antibodies to a major etiologic virus of human non-A, non-B hepatitis. Science 244:362–364Google Scholar
  25. 25.
    Lee W-H, Shew J-Y, Hong FD, Sery TW, Donoso LA, Young L-J, Bookstein R, Lee EY-HP (1987) The retinoblastoma susceptibility gene encodes a nuclear phosphoprote in associated with DNA binding activity. Nature 329:642–645Google Scholar
  26. 26.
    Liaw Y-F, Chu C-M, Lin D-Y, Sheen I-S, Yang C-Y, Huang M-J (1984) Age-specific prevalence and significance of hepatitis B e antigen and antibody in chronic hepatitis B virus infection in Taiwan: a comparison among asymptomatic carriers, chronic hepatitis, liver cirrhosis, and hepatocellular carcinoma. J Med Virol 13:385–391Google Scholar
  27. 27.
    Lin J-K, Chou C-K (1992) In vitro apoptosis in the human hepatoma cell line induced by transforming growth factor β 1. Cancer Res 52:385–388Google Scholar
  28. 28.
    Liu C, Tsao M-S, Grisham JW (1988) Transforming growth factors produced by normal and neoplastically transformed rat liver epithelial cells in culture. Cancer Res 48:850–855Google Scholar
  29. 29.
    Liu M-L, Mars WM, Zarnegar R, Michalopoulos GK (1994) Collagenase pretreatment and the mitogenic effects of hepatocyte growth factor and transforming growth factor-α in adult rat liver. Hepatology 19:1521–1527Google Scholar
  30. 30.
    Masaki S, Shiku H, Kaneda T, Koiwai O, Yoshida S (1982) Production and characterization of monoclonal antibody against IOS DNA polymerase α from calf thymus. Nucleic Acids Res 10:4703–4713Google Scholar
  31. 31.
    Mead JE, Fausto N (1989) Transforming growth factor a may be a physiological regulator of liver regeneration by means of an amocrine mechanism. Proc Natl Acad Sci USA 86:1558–1562Google Scholar
  32. 32.
    Mekori YA, Oh CK, Metcalfe DD (1993) IL-3-dependent murine mast cells undergo apoptosis on removal of IL-3: prevention of apoptosis byc-kit ligand. J Immunol 151:3775–3784Google Scholar
  33. 33.
    Mihara K, Cao X-R, Yen A, Chandler S, Driscoll B, Murphree AL, T'Ang A, Fung YKT (1989) Cell cycle-dependent regulation of phosphorylation of the human retinoblastoma gene product. Science 246:1300–1303Google Scholar
  34. 34.
    Nishioka K, Watanabe J, Furuta S, Tanaka E, Iino S, Suzuki H, Tsuji T, Yano M, Kuo G, Choo QL, Houghton M, Oda T (1991) A high prevalence of antibody to the hepatitis C virus in patients with hepatocellular carcinoma in Japan. Cancer 67:429–433Google Scholar
  35. 35.
    Oberhammer F, Bursch W, Tiefenbacher R, Froschl G, Pavelka M, Purchio T, Schulte-Hermann R (1993) Apoptosis is induced by transforming growth factor-β 1 within 5 hours in regressing liver without significant fragmentation of the DNA. Hepatology 18:1238–1246Google Scholar
  36. 36.
    Okamoto H, Kurai K, Okada S, Yamamoto K, Lizuka H, Tanaka T, Fukuda S, Tsuda F, Mishiro S (1992) Full-length sequence of a hepatitis C virus genome having poor homology to reported isolates: comparative study of four distinct genotypes. Virology 188:331–341Google Scholar
  37. 37.
    Okuda K, Liver Cancer Study Group of Japan (1980) Primary liver cancers in Japan. Cancer 45:2663–2669Google Scholar
  38. 38.
    Pittman RN, Wang S, DiBenedetto AJ, Mills JC (1993) A system for characterizing cellular and molecular events in programmed neuronal cell death. J Neurosci 13:3669–3680Google Scholar
  39. 39.
    Russel WE, Dempsey PJ, Sitaric S, Peck AJ, Coffey RJ Jr (1993) Transforming growth factor-α (TGF-α) concentrations increase in regenerating rat liver: evidence for a delayed accumulation of mature TGF-α. Endocrinology 133:1731–1738Google Scholar
  40. 40.
    Schaff Z, Hsia CC, Sarosi I, Tabor E (1994) Overexpression of transforming growth factor-α in hepatocellular carcinoma and focal nodular hyperplasia from European patients. Hum Pathol 25:644–651Google Scholar
  41. 41.
    Seki S, Kawakita N, Yanai A, Kitada T, Sakai Y, Nakatani K, Yamada T, Sakaguchi H, Kuroki T (1995) Expression of the retinoblastoma gene product in human hepatocellular carcinoma. Hum Pathol 26:366–374Google Scholar
  42. 42.
    Smith CA, Williams GT, Kingston R, Jenkinson EJ, Owen JJT (1989) Antibodies to CD3/T-cell receptor complex induce death by apoptosis in immature T cells in thymic cultures. Nature 337:181–184Google Scholar
  43. 43.
    Sporn MB, Roberts AB (1985) Autocrine growth factors and cancer. Nature 313:745–747Google Scholar
  44. 44.
    Steiner PE (1960) Cancer of the liver and cirrhosis in trans-Sahara Africa and the United States of America. Cancer 13:1085–1166Google Scholar
  45. 45.
    Tamano S, Merlino GT, Ward JM (1994) Rapid development of hepatic tumors in transforming growth factor α transgenic mice associated with increased cell proliferation in precancerous hepatocellular lesions initiated byN-nitrosodiethylamine and promoted by phenobarbital. Carcinogenesis 15:1791–1798Google Scholar
  46. 46.
    Tanaka K, Hirihata K, Koga S, Sugimachi K, Kanematsu T, Ohryohji F, Nawata H, Ishibashi H, Maeda Y, Kiyokawa H, Tokunaga K, Irita Y, Takashita S, Arase Y, Nishino N (1991) Hepatitis C and hepatitis B in the etiology of hepatocellular carcinoma in the Japanese population. Cancer Res 51:2842–2847Google Scholar
  47. 47.
    Vagas V, Castells L, Esteban JI (1990) High frequency of antibodies to the hepatitis C virus among patients with hepatocellular carcinoma. Ann Intern Med 112:232–233Google Scholar
  48. 48.
    Webber EM, FitzGerald MJ, Brown PI, Barlett MH, Fausto N (1993) Transforming growth factor-α expression during liver regeneration after partial hepatectomy and toxic injury, and hepatocyte growth factor. Hepatology 18:1422–1431Google Scholar
  49. 49.
    Webber EM, Godowski PJ, Fausto N (1994) In vivo response of hepatocytes to growth factors requires an initial priming stimulus. Hepatology 19:489–497Google Scholar
  50. 50.
    Wrong ST, Winchell LF, McCune BK, Earp HS, Teixido J, Massague J, Herman B, Lee DC (1989) The TGF-α precursor expressed on the cell surface binds to the EGF receptor on adjacent cells, leading to signal transduction. Cell 56:495–506Google Scholar
  51. 51.
    Wyllie AH (1980) Glucocorticoid-induced thymocyte apoptosis is associated with endogenous endonuclease activation. Nature 284:555–556Google Scholar
  52. 52.
    Wyllie AH, Kerr JFR, Currie AR (1980) Cell death: the significance of apoptosis. Int Rev Cytol 68:251–306Google Scholar

Copyright information

© Springer-Verlag 1997

Authors and Affiliations

  • Shuichi Seki
    • 1
  • Yasuhiko Sakai
    • 1
  • Takuya Kitada
    • 1
  • Nobuyoshi Kawakita
    • 1
  • Atsushi Yanai
    • 1
  • Hiroko Tsutsui
    • 2
  • Hiroki Sakaguchi
    • 1
  • Tetsuo Kuroki
    • 1
  • Takeyuki Monna
    • 3
  1. 1.Third Department of Internal MedicineOsaka City University Medical SchoolOsakaJapan
  2. 2.Third Department of BiochemistryOsaka City University Medical SchoolOsakaJapan
  3. 3.Department of Public HealthOsaka City University Medical SchoolOsakaJapan

Personalised recommendations