Induction of hsp 70 in HepG2 cells in response to hepatotoxicants.
The objective of this study was to determine if a variety of hepatotoxicants could induce the level of heat shock protein 70I, and whether or not elevated levels of heat shock proteins (hsp's) could provide cytoprotection from those hepatotoxicants. Exposure of HepG2 cells to cytotoxic concentrations of bromobenzene, cadmium, cyclophosphamide, or diethylnitrosamine increased the level of hsp 70I protein and mRNA, while carbon tetrachloride and cocaine had no effect on hsp 70I or mRNA levels. To determine if induction of hsp 70I might afford protection against cytotoxicity, HepG2 cells were given a prior sublethal heat shock (sub-LHS) (43 degrees C for 1 hr) to induce hsp's and then challenged 24 hr later with the hepatotoxicants. Sub-LHS pretreatment diminished toxicity from bromobenzene, cadmium, cyclophosphamide, or diethyl-nitrosamine, but not carbon tetrachloride or cocaine. In cells treated with [14C]carbon tetrachloride or [3H]cocaine, no detectable covalent binding to proteins was observed; whereas, [14C]-bromobenzene treatment resulted in substantial covalent binding to cellular protein. The apparent absence of formation of reactive metabolite adducted proteins from cocaine and carbon tetrachloride may explain why no hsp 70I induction was observed with these agents. The correlation between hepatotoxicant induction of hsp 70I and cytoprotection afforded by sub-LHS pretreatment suggests that hsp 70I induction may represent an important cellular defense mechanism in the liver.
Salminen
W F
WF
Department of Pharmacology and Therapeutics, J. Hillis Miller Health Science Center, University of Florida, Gainesville 32601, USA.
Voellmy
R
R
Roberts
S M
SM
D001969:Bromobenzenes / Q000378:metabolism / Q000633:toxicity*; D002104:Cadmium / Q000633:toxicity*; D002251:Carbon Tetrachloride / Q000378:metabolism; D002273:Carcinogens / Q000633:toxicity*; D002478:Cells, Cultured; D002849:Chromatography, Gas; D003042:Cocaine / Q000378:metabolism; D003520:Cyclophosphamide / Q000633:toxicity*; D004052:Diethylnitrosamine / Q000633:toxicity*; D004591:Electrophoresis, Polyacrylamide Gel; D018840:HSP70 Heat-Shock Proteins / Q000378:metabolism*; D018869:Heat-Shock Response / Q000187:drug effects*; D006801:Humans; D008113:Liver Neoplasms / Q000378:metabolism*
D001969:Bromobenzenes; D002273:Carcinogens; D018840:HSP70 Heat-Shock Proteins; D002104:Cadmium; D004052:Diethylnitrosamine; D003520:Cyclophosphamide; D002251:Carbon Tetrachloride; C032036:bromobenzene; D003042:Cocaine