Effect of phenobarbital and 3-methylcholanthrene on aldehyde dehydrogenase activity in cultures of HepG2 cells and normal human hepatocytes

https://doi.org/10.1016/0009-2797(87)90080-9Get rights and content

Abstract

Aldehyde dehydrogenase (ALDH) activity was measured in primary cultures of normal human hepatocytes and of the human hepatoma cell line HepG2 after application of phenobarbital (PB) or 3-methylcholanthrene (MC) for 5 days. Treatment with PB alone resulted in a significant increase in both protein and DNA content at concentrations of 2 and 3 mM. Treatment with MC at a concentration as low as 5 μM led to a significant loss of cells when it lasted more than 5 days. Concentrations of 3–5 mM of PB in the media of HepG2 cell cultures caused a 2-fold enhancement of the activity of ALDH, as measured with NAD and propionaldehyde (P/NAD) or benzaldehyde (B/NAD). On the other hand, MC-treated cultures (5 μM) showed a 20-fold increase in enzyme activity measured with NADP and benzaldehyde (B/NADP), and a 2-fold increase in B/NAD activity. Combined treatment with both PB and MC led to an effect of dynamic synergism as far as B/NAD and B/NADP activities are concerned, suggesting a metabolite of MC as the mediator for the increase of ALDH activity. Normal human hepatocytes in primary cultures responded to PB (3 mM) in a similar way as HepG2 cells as far as DNA and protein content and ALDH activity are concerned. It is concluded, that HepG2 hepatoma cells behave similar to the normal hepatocytes in terms of ALDH regulation and can be used for studies on the activity of ALDH as modified by added xenobiotics.

References (50)

  • R. Lindahl et al.

    Characterization of aldehyde dehydrogenase from HTC rat hepatoma cells

    Biochim. Biophys. Acta

    (1985)
  • S.C. Strom et al.

    Collagen as a substrate for cell growth and differentiation

    Methods Enzymol.

    (1982)
  • M.M. Bradford

    A rapid and sensitive method for the quantitation of microgram quantities of protein, utilizing the principle of protein-dye binding

    Anal. Biochem.

    (1976)
  • R.T. Hinegardner

    An improved fluorometric assay for DNA

    Anal. Biochem.

    (1971)
  • J.M. Rash et al.

    Lipoprotein apolipoprotein synthesis by human hepatoma cells in culture

    Biochim. Biophys. Acta

    (1981)
  • S.-P. Tam et al.

    Effects of estrogen on apolipoprotein secretion by the human hepatocarcinoma cell line HepG2

    J. Biol. Chem.

    (1985)
  • K.L. Dearfield et al.

    Evaluation of a human hepatoma cell line as a target cell in genetic toxicology

    Mutat. Res.

    (1983)
  • K.L. Dearfield et al.

    In vitro assays of in vivo exposure to cyclophosphamide: Induction of sister-chromatid exchanges in peripheral lymphocytes, bone-marrow cells and in cultured cells exposed to plasma

    Mutat. Res.

    (1985)
  • J.R. Dawson et al.

    Induction of drug-metabolising enzymes in human liver cell line HepG2

    FEBS Lett.

    (1985)
  • M. Miyazaki et al.

    Long-term survival of functional hepatocytes from adult rat in the presence of phenobarbital in primary culture

    Exp. Cell Res.

    (1985)
  • A.M. Edwards et al.

    Phenobarbital and some other liver tumor promoters stimulate DNA synthesis in cultured rat hepatocytes

    Biochem. Biophys. Res. Commun.

    (1985)
  • A.H. Blair et al.

    Human liver aldehyde dehydrogenase: partial purification and properties

    Can. J. Biochem.

    (1969)
  • T. Koivula

    Subcellular distribution and characterization of human liver adehyde dehydrogenase fractions

    Life Sci.

    (1975)
  • S. Harada et al.

    Isozyme variations in acetaldehyde dehydrogenase (E.C. 1.2.1.3) in human tissues

    Hum. Genet.

    (1978)
  • R. Pietruszko et al.

    Chemical modification and site of interaction of human aldehyde dehydrogenase E1 with disulfiram and iodoacetamide

  • Cited by (32)

    • Effects of 3-methylcholanthrene and aspirin co-administration on ALDH3A1 in HepG2 cells

      2001, Chemico-Biological Interactions
      Citation Excerpt :

      Induction of the enzymes mentioned above by carcinogenic substances has also been described in these cells [10,33]. Our data confirm previous findings on time and dose response in the induction of ALDH3A1 activity after exposure of HepG2 cells to 3MC [10]. From these observations, it can be concluded that the concentrations of 3MC needed to enhance the ALDH3A1 activity are rather toxic for the cells.

    View all citing articles on Scopus
    View full text