A comparison of apoptosis and necrosis induced by hepatotoxins in HepG2 cells.
7H-Dibenzo[c,g]carbazole (DBC), an N-heterocyclic aromatic hydrocarbon, is cytotoxic and carcinogenic in rodent liver. While DBC leads to necrotic lesions in the liver, the induction of apoptosis by DBC has not been investigated. The focus of this study was to determine the degree to which apoptosis and necrosis contributed to DBC cytotoxicity in a human hepatoma cell line (HepG2). To determine if these effects were unique to DBC, the results were compared to another hepatotoxin, aflatoxin B(1) (AFB(1)). DBC produced a distinct biphasic LDH release curve within 24 h of exposure. During the same time period lower concentrations of DBC (<10 microM) induced the formation of DBC-DNA adducts and increased p53 protein levels followed by apoptotic cell death. However, increasing the concentration of DBC to 80 microM led to lower DNA adduct and p53 protein levels. At this concentration, intracellular ATP levels were rapidly depleted followed by cell swelling and loss of membrane integrity consistent with necrotic cell death. In contrast to DBC, a biphasic LDH release curve was not observed for AFB(1). Instead, AFB(1) induced a concentration-dependent increase in apoptosis that reached two- to threefold higher levels than DBC. These results suggest that differences exist in the extent and type of cell death induced by DBC and AFB(1) at equimolar concentrations. Apoptosis and necrosis result from low and high concentrations of DBC, respectively, and may be dependent upon intracellular ATP levels.
O'Brien
T
T
Department of Environmental Health, University of Cincinnati Medical Center, Cincinnati, Ohio, 45267-0056, USA.
Babcock
G
G
Cornelius
J
J
Dingeldein
M
M
Talaska
G
G
Warshawsky
D
D
Mitchell
K
K
D000255:Adenosine Triphosphate / Q000378:metabolism; D016604:Aflatoxin B1 / Q000494:pharmacology; D017209:Apoptosis; D002227:Carbazoles / Q000494:pharmacology*; D002273:Carcinogens / Q000494:pharmacology*; D004249:DNA Damage / Q000187:drug effects; D053938:DNA Fragmentation; D018197:Hepatoblastoma / Q000473:pathology*; D006801:Humans; D020287:In Situ Nick-End Labeling; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008113:Liver Neoplasms / Q000139:chemically induced / Q000473:pathology*; D008854:Microscopy, Electron; D009336:Necrosis; D014407:Tumor Cells, Cultured; D016159:Tumor Suppressor Protein p53 / Q000378:metabolism
D002227:Carbazoles; D002273:Carcinogens; D016159:Tumor Suppressor Protein p53; D000255:Adenosine Triphosphate; D016604:Aflatoxin B1; D007770:L-Lactate Dehydrogenase; C013478:7H-dibenzo(c,g)carbazole