Inhibition of benzo[a]pyrene metabolism by insulin, FITC-insulin and an FITC-insulin-antibody conjugate in the human hepatoma cell line HepG2.
Benzo[a]pyrene (BaP) can be metabolically activated to an ultimate carcinogen, (+)-anti-BaP-7,8-dihydrodiol-9,10-epoxide [(+)-anti-BaPDE] by cells in culture. This activation involves oxidation by specific isoforms of cytochrome P450s such as CYP1A1. The human hepatoma cell line, HepG2, was used to examine the effect of inhibition of CYP1A1 activity by anti CYP1A1 specific antibodies on BaP metabolism. Metabolism of BaP to water-soluble metabolites by HepG2 cells in culture was 50% lower in fluorescein isothiocyanate (FITC)-insulin-CYP1A1-antibody-conjugate-treated cells than in control cells. However, FITC-insulin (lacking anti CYP1A1 conjugates) or insulin alone also decreased BaP metabolism by 50%. This insulin-induced inhibition of BaP metabolism was observed for cultures treated with a concentration range of FITC-insulin from 50-1000 nM. FITC-conjugated gamma-globulin showed no significant binding to HepG2 cells by fluorescence microscopy, however, FITC-insulin-antibody conjugates bound extensively, suggesting that FITC-insulin conjugates still retain the ability to bind insulin receptors. These results demonstrate that free insulin, FITC-insulin or FITC-insulin conjugated to antibodies are effective inhibitors of BaP metabolism in cells in culture.
Polzer
R J
RJ
Department of Medicinal Chemistry and Pharmacognosy, Purdue University, West Lafayette, IN 47907, USA.
Coffing
S L
SL
Marcus
C B
CB
Park
S S
SS
Gelboin
H V
HV
Baird
W M
WM
D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D001564:Benzo(a)pyrene / Q000378:metabolism*; D006528:Carcinoma, Hepatocellular; D065607:Cytochrome P-450 Enzyme Inhibitors; D003577:Cytochrome P-450 Enzyme System / Q000276:immunology / Q000378:metabolism; D016650:Fluorescein-5-isothiocyanate / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology; D006801:Humans; D007328:Insulin / Q000031:analogs & derivatives* / Q000378:metabolism / Q000494:pharmacology*; D011972:Receptor, Insulin / Q000378:metabolism; D014407:Tumor Cells, Cultured
D000906:Antibodies; D065607:Cytochrome P-450 Enzyme Inhibitors; D007328:Insulin; C031573:insulin, fluorescein-isothiocyanated-; D001564:Benzo(a)pyrene; D003577:Cytochrome P-450 Enzyme System; D011972:Receptor, Insulin; D016650:Fluorescein-5-isothiocyanate