Skip to main content
PMC home page
The Journal of Cell Biology logoLink to The Journal of Cell Biology
. 1994 Apr 1;125(1):197–203. doi: 10.1083/jcb.125.1.197

Suppression of liver cell apoptosis in vitro by the non-genotoxic hepatocarcinogen and peroxisome proliferator nafenopin

PMCID: PMC2120009  PMID: 8138571

Abstract

Suppression of apoptosis has been implicated as a mechanism for the hepatocarcinogenicity of the peroxisome proliferator class of non- genotoxic carcinogens. The ability of the peroxisome proliferator nafenopin to suppress or delay the onset of liver apoptosis was investigated using primary cultures of rat hepatocytes and the Reuber hepatoma cell line FaO. 50 microM nafenopin reversibly maintained the viability of primary rat hepatocyte cultures which otherwise degenerated within 8 d of establishment. The maintenance of viability of hepatocyte monolayers was associated with a significant decrease in the number of cells exhibiting chromatin condensation patterns typical of apoptosis. Apoptosis could be induced in hepatocytes by administration of 5 ng/ml TGF beta 1. Co-addition of 50 microM nafenopin significantly reduced TGF beta 1-induced apoptosis by 50-60%. TGF beta 1 (1-5 ng/ml) also induced apoptosis in the FaO rat hepatoma cell line. Cell death was accompanied by detachment of FaO cells from the monolayer and detached cells exhibited chromatin condensation and non-random DNA fragmentation patterns typical of apoptosis. Co-addition of 50 microM nafenopin to TGF beta 1-treated FaO cultures significantly reduced the number of apoptotic cells detaching from the monolayer at 24 h. In contrast, nafenopin had no significant effect on FaO apoptosis induced by the DNA damaging agents etoposide and hydroxyurea. We conclude that suppression of liver cell death by apoptosis may play a role in the hepatocarcinogenicity of the peroxisome proliferators, although the extent of this protection is dependent on the nature of the apoptotic stimulus.

Full Text

The Full Text of this article is available as a PDF (1.4 MB).

Selected References

These references are in PubMed. This may not be the complete list of references from this article.

  1. Bayly A. C., French N. J., Dive C., Roberts R. A. Non-genotoxic hepatocarcinogenesis in vitro: the FaO hepatoma line responds to peroxisome proliferators and retains the ability to undergo apoptosis. J Cell Sci. 1993 Feb;104(Pt 2):307–315. doi: 10.1242/jcs.104.2.307. [DOI] [PubMed] [Google Scholar]
  2. Braun L., Gruppuso P., Mikumo R., Fausto N. Transforming growth factor beta 1 in liver carcinogenesis: messenger RNA expression and growth effects. Cell Growth Differ. 1990 Mar;1(3):103–111. [PubMed] [Google Scholar]
  3. Bursch W., Düsterberg B., Schulte-Hermann R. Growth, regression and cell death in rat liver as related to tissue levels of the hepatomitogen cyproterone acetate. Arch Toxicol. 1986 Dec;59(4):221–227. doi: 10.1007/BF00290542. [DOI] [PubMed] [Google Scholar]
  4. Bursch W., Lauer B., Timmermann-Trosiener I., Barthel G., Schuppler J., Schulte-Hermann R. Controlled death (apoptosis) of normal and putative preneoplastic cells in rat liver following withdrawal of tumor promoters. Carcinogenesis. 1984 Apr;5(4):453–458. doi: 10.1093/carcin/5.4.453. [DOI] [PubMed] [Google Scholar]
  5. Bursch W., Oberhammer F., Jirtle R. L., Askari M., Sedivy R., Grasl-Kraupp B., Purchio A. F., Schulte-Hermann R. Transforming growth factor-beta 1 as a signal for induction of cell death by apoptosis. Br J Cancer. 1993 Mar;67(3):531–536. doi: 10.1038/bjc.1993.98. [DOI] [PMC free article] [PubMed] [Google Scholar]
  6. Bursch W., Oberhammer F., Schulte-Hermann R. Cell death by apoptosis and its protective role against disease. Trends Pharmacol Sci. 1992 Jun;13(6):245–251. doi: 10.1016/0165-6147(92)90077-j. [DOI] [PubMed] [Google Scholar]
  7. Cattley R. C., Richardson K. K., Smith-Oliver T., Popp J. A., Butterworth B. E. Effect of peroxisome proliferator carcinogens on unscheduled DNA synthesis in rat hepatocytes determined by autoradiography. Cancer Lett. 1986 Dec;33(3):269–277. doi: 10.1016/0304-3835(86)90066-2. [DOI] [PubMed] [Google Scholar]
  8. Dreyer C., Keller H., Mahfoudi A., Laudet V., Krey G., Wahli W. Positive regulation of the peroxisomal beta-oxidation pathway by fatty acids through activation of peroxisome proliferator-activated receptors (PPAR). Biol Cell. 1993;77(1):67–76. doi: 10.1016/s0248-4900(05)80176-5. [DOI] [PubMed] [Google Scholar]
  9. Issemann I., Green S. Activation of a member of the steroid hormone receptor superfamily by peroxisome proliferators. Nature. 1990 Oct 18;347(6294):645–650. doi: 10.1038/347645a0. [DOI] [PubMed] [Google Scholar]
  10. Lazarow P. B., De Duve C. A fatty acyl-CoA oxidizing system in rat liver peroxisomes; enhancement by clofibrate, a hypolipidemic drug. Proc Natl Acad Sci U S A. 1976 Jun;73(6):2043–2046. doi: 10.1073/pnas.73.6.2043. [DOI] [PMC free article] [PubMed] [Google Scholar]
  11. Lin J. K., Chou C. K. In vitro apoptosis in the human hepatoma cell line induced by transforming growth factor beta 1. Cancer Res. 1992 Jan 15;52(2):385–388. [PubMed] [Google Scholar]
  12. Marsman D. S., Cattley R. C., Conway J. G., Popp J. A. Relationship of hepatic peroxisome proliferation and replicative DNA synthesis to the hepatocarcinogenicity of the peroxisome proliferators di(2-ethylhexyl)phthalate and [4-chloro-6-(2,3-xylidino)-2-pyrimidinylthio]acetic acid (Wy-14,643) in rats. Cancer Res. 1988 Dec 1;48(23):6739–6744. [PubMed] [Google Scholar]
  13. Moody D. E., Gibson G. G., Grant D. F., Magdalou J., Rao M. S. Peroxisome proliferators, a unique set of drug-metabolizing enzyme inducers: commentary on a symposium. Drug Metab Dispos. 1992 Nov-Dec;20(6):779–791. [PubMed] [Google Scholar]
  14. Moody D. E., Reddy J. K., Lake B. G., Popp J. A., Reese D. H. Peroxisome proliferation and nongenotoxic carcinogenesis: commentary on a symposium. Fundam Appl Toxicol. 1991 Feb;16(2):233–248. doi: 10.1016/0272-0590(91)90108-g. [DOI] [PubMed] [Google Scholar]
  15. Oberhammer F. A., Pavelka M., Sharma S., Tiefenbacher R., Purchio A. F., Bursch W., Schulte-Hermann R. Induction of apoptosis in cultured hepatocytes and in regressing liver by transforming growth factor beta 1. Proc Natl Acad Sci U S A. 1992 Jun 15;89(12):5408–5412. doi: 10.1073/pnas.89.12.5408. [DOI] [PMC free article] [PubMed] [Google Scholar]
  16. Orton T. C., Parker G. L. The effect of hypolipidemic agents on the hepatic microsomal drug-metabolizing enzyme system of the rat. Induction of cytochrome(s) P-450 with specificity toward terminal hydroxylation of lauric acid. Drug Metab Dispos. 1982 Mar-Apr;10(2):110–115. [PubMed] [Google Scholar]
  17. Rao M. S., Reddy J. K. An overview of peroxisome proliferator-induced hepatocarcinogenesis. Environ Health Perspect. 1991 Jun;93:205–209. doi: 10.1289/ehp.9193205. [DOI] [PMC free article] [PubMed] [Google Scholar]
  18. Reddy J. K., Lalwai N. D. Carcinogenesis by hepatic peroxisome proliferators: evaluation of the risk of hypolipidemic drugs and industrial plasticizers to humans. Crit Rev Toxicol. 1983;12(1):1–58. doi: 10.3109/10408448309029317. [DOI] [PubMed] [Google Scholar]
  19. Reddy J. K., Rao M. S. Malignant tumors in rats fed nafenopin, a hepatic peroxisome proliferator. J Natl Cancer Inst. 1977 Dec;59(6):1645–1650. doi: 10.1093/jnci/59.6.1645. [DOI] [PubMed] [Google Scholar]
  20. Reddy J. K., Rao M. S. Oxidative DNA damage caused by persistent peroxisome proliferation: its role in hepatocarcinogenesis. Mutat Res. 1989 Sep;214(1):63–68. doi: 10.1016/0027-5107(89)90198-x. [DOI] [PubMed] [Google Scholar]
  21. Richardson U. I., Tashjian A. H., Jr, Levine L. Establishment of a clonal strain of hepatoma cells which secrete albumin. J Cell Biol. 1969 Jan;40(1):236–247. doi: 10.1083/jcb.40.1.236. [DOI] [PMC free article] [PubMed] [Google Scholar]
  22. Schulte-Hermann R. Induction of liver growth by xenobiotic compounds and other stimuli. CRC Crit Rev Toxicol. 1974 Sep;3(1):97–158. doi: 10.3109/10408447409079856. [DOI] [PubMed] [Google Scholar]
  23. Seglen P. O. Preparation of rat liver cells. 3. Enzymatic requirements for tissue dispersion. Exp Cell Res. 1973 Dec;82(2):391–398. doi: 10.1016/0014-4827(73)90357-1. [DOI] [PubMed] [Google Scholar]
  24. Sher T., Yi H. F., McBride O. W., Gonzalez F. J. cDNA cloning, chromosomal mapping, and functional characterization of the human peroxisome proliferator activated receptor. Biochemistry. 1993 Jun 1;32(21):5598–5604. doi: 10.1021/bi00072a015. [DOI] [PubMed] [Google Scholar]
  25. Warren J. R., Simmon V. F., Reddy J. K. Properties of hypolipidemic peroxisome proliferators in the lymphocyte [3H]thymidine and Salmonella mutagenesis assays. Cancer Res. 1980 Jan;40(1):36–41. [PubMed] [Google Scholar]
  26. Yanagihara K., Tsumuraya M. Transforming growth factor beta 1 induces apoptotic cell death in cultured human gastric carcinoma cells. Cancer Res. 1992 Jul 15;52(14):4042–4045. [PubMed] [Google Scholar]

Articles from The Journal of Cell Biology are provided here courtesy of The Rockefeller University Press

RESOURCES