Stable transfection of LLC-PK1 cells with human microsomal glutathione S-transferase gene increases haloalkene glutathione S-conjugate formation and cytotoxicity.
Nephrotoxic haloalkenes undergo glutathione- and cysteine conjugate beta-lyase-dependent bioactivation, and glutathione S-conjugate formation with haloalkenes as substrates is preferentially catalyzed by the hepatic microsomal glutathione S-transferase (mGST). Porcine kidney-derived LLC-PK1 cells, which are competent to bioactivate glutathione and cysteine S-conjugates of haloalkenes, show low mGST activity. Stable transfection of LLC-PK1 cells with the gene encoding mGST would be expected to increase glutathione S-conjugate formation and, therefore, to increase haloalkene cytotoxicity. Transfection of LLC-PK1 cells with human mGST genes resulted in increased expression of mGST protein in microsomal fractions, in increased glutathione S-conjugate formation with hexachloro-1,3-butadiene and 1-chloro-2,4-dinitrobenzene as the substrates, and in increased cytotoxicity of hexachloro-1,3-butadiene. In addition, transfection with mGST gene also increased the activity of cytosolic glutathione S-transferases.
Otieno
M A
MA
Department of Pharmacology and Physiology, University of Rochester, School of Medicine and Dentistry, New York 14642, USA.
Anders
M W
MW
D000475:Alkenes / Q000378:metabolism* / Q000493:pharmacokinetics / Q000633:toxicity*; D000818:Animals; D001711:Biotransformation; D003545:Cysteine / Q000378:metabolism; D003600:Cytosol / Q000378:metabolism; D003603:Cytotoxins / Q000378:metabolism* / Q000493:pharmacokinetics / Q000633:toxicity*; D005978:Glutathione / Q000031:analogs & derivatives* / Q000378:metabolism; D005982:Glutathione Transferase / Q000145:classification / Q000235:genetics* / Q000378:metabolism*; D006801:Humans; D007150:Immunohistochemistry; D007668:Kidney / Q000187:drug effects / Q000378:metabolism; D018374:LLC-PK1 Cells; D008861:Microsomes / Q000378:metabolism; D013552:Swine; D014162:Transfection
D000475:Alkenes; D003603:Cytotoxins; D005982:Glutathione Transferase; D005978:Glutathione; D003545:Cysteine