gamma-Glutamyltranspeptidase-positive rat hepatocytes are protected from GSH depletion, oxidative stress and reversible alterations of collagen receptors.
The aim of this study has been to define cytotoxic mechanisms that may cause clonal expansion in the liver of pre-carcinogenic cells. An in vitro model, which has been described previously, was used. Hepatocytes were isolated from carcinogen-treated rats and a high proportion of the cells were gamma-glutamyltranspeptidase (GGT)-positive. The cells were incubated in suspension and exposed to toxic agents in concentrations that induced a moderate increase in cellular leakage within 3 h. Samples were withdrawn and sampled cells were then allowed to attach to collagen-coated plates. Attached cells were stained and the ratio of GGT-positive/GGT-negative cells (GGT-ratio) was determined. The initial GGT-ratio was 10.4 +/- 4.7% and an increased ratio was taken as a sign of toxicity that resulted in a selection of GGT-positive cells. In a first series of experiments it was shown that hydroquinone and menadione increase the GGT-ratio, while diquat, sodium selenite, diethyl maleate or phorone do not. However, diethyl maleate in combination with diquat increased the GGT-ratio. Hydrogen peroxide (5 mM) increased the GGT-ratio as effectively as hydroquinone (0.3 mM). Lower concentrations of H2O2 (0.05 mM) increased the GGT-ratio in GSH-depleted cells. The changes induced by hydroquinone and H2O2 in low concentration were reversible. In another series of experiments, plates coated with antibodies against beta 1-integrin were used. An increase in the GGT-ratio was obtained with anti beta 1-integrin, but not with broad spectrum anti-rat hepatocyte or anti-rat beta 2-microglobulin antibodies as substrata. These data suggested an involvement of the beta 1-integrin in the selection. Taken together, these data indicate that GGT-positive hepatocytes are protected against GSH depletion and oxidative stress that may result in reversible receptor alterations.
Stenius
U
U
Department of Toxicology, Institute of Occupational Health, 84 Solna, Sweden.
Rubin
K
K
Gullberg
D
D
Högberg
J
J
D000818:Animals; D014408:Biomarkers, Tumor / Q000032:analysis*; D002471:Cell Transformation, Neoplastic; D002478:Cells, Cultured; D003094:Collagen / Q000378:metabolism; D004052:Diethylnitrosamine / Q000633:toxicity; D004178:Diquat / Q000494:pharmacology; D005260:Female; D005978:Glutathione / Q000378:metabolism*; D006861:Hydrogen Peroxide / Q000494:pharmacology*; D006873:Hydroquinones / Q000494:pharmacology; D007659:Ketones / Q000494:pharmacology; D007700:Kinetics; D008099:Liver / Q000187:drug effects / Q000201:enzymology* / Q000378:metabolism; D008298:Maleates / Q000494:pharmacology; D010634:Phenobarbital / Q000494:pharmacology; D051381:Rats; D011956:Receptors, Cell Surface / Q000187:drug effects / Q000378:metabolism*; D038981:Receptors, Collagen; D012643:Selenium / Q000494:pharmacology; D018038:Sodium Selenite; D014812:Vitamin K / Q000494:pharmacology; D005723:gamma-Glutamyltransferase / Q000032:analysis*
D014408:Biomarkers, Tumor; D006873:Hydroquinones; D007659:Ketones; D008298:Maleates; D011956:Receptors, Cell Surface; D038981:Receptors, Collagen; D014812:Vitamin K; D004052:Diethylnitrosamine; C018637:phorone; D003094:Collagen; D004178:Diquat; D006861:Hydrogen Peroxide; D005723:gamma-Glutamyltransferase; C014476:diethyl maleate; D005978:Glutathione; D012643:Selenium; D018038:Sodium Selenite; D010634:Phenobarbital