Protective activity of different hepatic cytosolic glutathione S-transferases against DNA-binding metabolites of aflatoxin B1.

To evaluate the role of glutathione S-transferase (GST) isoenzymes in induced resistance of hepatocytes to aflatoxin B1 (AFB1), we compared DNA protective activities of different hepatic cytosol preparations and purified GSTs from normal rats, rats exposed to different polychlorinated biphenyls (PCBs), and rats with carcinogen-induced hepatocellular neoplasms, with cytosols or purified GSTs from mouse, rainbow trout, and human livers. These comparisons were performed in an in vitro assay for [3H]AFB1-DNA binding after activation by rat liver microsomes. Cytosol and S-hexylglutathione-affinity-purified GST preparations from livers of mice consistently had strong protective activity against AFB1-DNA binding. The majority of this activity was dependent on the presence of reduced glutathione (GSH) but some GSH-independent protection was observed in mouse hepatic cytosol, but not in purified GST preparations. We found that all of the GSH-dependent DNA-protective activity in mouse liver eluted as a single GST isoenzyme by hydroxyapatite chromatography. Preparations of cytosol and purified GSTs from normal rat liver, rainbow trout liver, and human liver had much less AFB1-specific DNA protective activity than GSTs found in mouse liver preparations. Cytosol from rats with carcinogen-generated liver neoplasms and livers induced with 3,3',4,4'-tetrachlorobiphenyl and 2,2',4,4',5,5'-hexachlorobiphenyl had more GST activity toward CDNB than cytosol from normal rat liver. When equivalent units of GST activity (CDNB) were compared, there was little difference observed between the DNA-protective activities of PCB-induced and normal rat liver cytosols, yet cytosol from rat liver neoplasms was more protective. Purified GST-P (7-7), the GST isoenzyme most induced in carcinogen-generated rat liver neoplasms, was not protective when added at protein concentrations found to be protective for total GSTs isolated from these neoplasms. These studies demonstrate that the resistance of mouse liver to AFB1 can be explained primarily by a single constitutive GST isoenzyme (YaYa or 4-4) with a relatively high activity toward DNA-binding metabolites of AFB1. GST isoenzymes with such high specific DNA protective activity against AFB1 metabolites were not evident in human, rat, or rainbow trout liver or in PCB-induced or neoplastic rat liver preparations.

Pathogenesis of skin and liver neoplasms in white suckers from industrially polluted areas in Lake Ontario.

Increased prevalences of epidermal and hepatobiliary neoplasms in white suckers (Catostomu commersoni) and brown bullheads (Ictalurus nebulosus) in the Western region of Lake Ontario have been associated with industrial pollution, but the identity and causative role of environmental carcinogens have not yet been established. Most epidermal tumors of lip and body skin are benign focal proliferations that occur in fish from the polluted Hamilton region, and also in fish from less polluted sites in the Great Lakes. These skin tumors in white suckers do not have consistent alterations in cellular glutathione S-transferases (GST), suggesting that growth of skin tumors is not promoted by chemicals normally detoxified by GST. However, elevated levels of glutathione peroxidase (GPO) and glutathione reductase (GR) in skin papillomas are indicative of promotional peroxidative tissue injury, either caused directly by xenobiotics or indirectly by chemical-induced inflammation. Liver tumors in white suckers from Lake Ontario include preneoplastic, benign, and malignant populations of hepatocellular and biliary cells, all of which are more prevalent in fish from polluted sites. These liver tumors are consistently associated with chronic cholangiohepatitis and segmental cholangiofibrosis, but these conditions also occur in white suckers in non-industrial locations. Thus, the natural occurrence of biliary disease, not attributable to industrial pollution, may have some influence on the development of liver tumors. Some preneoplastic lesions and the majority of neoplastic hepatocellular and biliary lesions in white suckers have low levels of total GST, indicating that these liver neoplasms are not promoted by xenobiotics normally detoxified by hepatic GSTs.(ABSTRACT TRUNCATED AT 250 WORDS)

Degradation of extracellular thymidine by cultured hepatocytes from rainbow trout (Salmo gairdneri).

1. Trout hepatocytes cultured as attached monolayers had low rates of [3H]-thymidine ([3H]-TdR) incorporation during replicative or repair synthesis of DNA. 2. Within 2 hr, most [3H]-TdR was metabolized by trout hepatocytes to a major product that eluted in advance of intact [3H]-TdR on Sephacryl S-200 columns. 3. Metabolism of [3H]-TdR by trout hepatocytes rapidly destroyed its ability to label replicating indicator cultures of proliferating rat hepatocytes. 4. These studies demonstrate that [3H]-TdR tracer assays for DNA synthesis cannot be reliably used in cultured trout hepatocytes which catabolize thymidine much more rapidly than do rat hepatocytes.