In vitro metabolism of aflatoxin B1 by normal and tumorous liver tissue from Thailand.
Liver tissues were obtained from 20 liver cancer patients from Thailand, an area where the incidence of this tumour is high and where exposure to aflatoxin occurs. The expression of hepatic cytochrome P450s (P450) and glutathione S-transferase (GST) was examined and this expression was compared to the in vitro metabolism of aflatoxin B1 (AFB1). There was a > 10-fold inter-individual variation in expression of the various P450s including CYP3A4 (57-fold), CYP2B6 (56-fold) and CYP2A6 (120-fold). Microsomal metabolism of AFB1 to AFB1 8,9-epoxide (as measured by AFB1 tris-diol formation) and aflatoxin Q1 (AFQ1), the major metabolite produced, was significantly correlated with CYP3A3/4 expression (P < 0.001) and, to a lesser extent, with CYP2B6 expression (P < 0.01). There was a significantly reduced expression of major P450 proteins in microsomes from liver tumours compared to microsomes from the paired normal liver when analysed by Western immunoblot analysis. The production of AFQ1 and AFB1 tris-diol was almost uniformly reduced in tumours, but interestingly, the production of AFP1 was significantly increased. The immunoreactive expression of the major human classes of cytosolic GSTs (alpha, mu and pi) was also analyzed in normal and tumorous liver tissue. The expression of GSTA (alpha) and GSTM (mu) class proteins was markedly decreased and GSTP (pi) increased in the majority of tumour cytosols compared to normal liver. The cytosolic GST activity (1-chloro-2,4-dinitrobenzene conjugation) was significantly lower in liver tumours compared to normal liver (193 +/- 149 versus 875 +/- 299 nmol/min/mg, P < 0.0001), as was glutathione peroxidase (GPx) activity (cumene hydroperoxide) (26 +/- 23 versus 70 +/- 26 nmol/min/mg respectively, P < 0.005). Ten out of 14 individuals (71%) were homozygous null when genotyped for GSTM1. There was no detectable conjugation of AFB1 8,9-epoxide to glutathione by cytosol either from tumorous or normal liver. Thus, capacity of human cytosols to conjugate reactive AFB1 metabolites to GSH resembled AFB1-sensitive species such as rat, trout and duck rather than resistant species such as mouse and hamster. These data indicate a strong capacity of multiple forms of human hepatic P450s to metabolize AFB1 to both the reactive intermediate AFB1 8,9-epoxide and the detoxification product AFQ1. These results suggest that in view of the lack of significant GST-mediated protection against AFB1 in human liver, variations in expression of hepatic P450, due either to genetic polymorphisms or to modulation by environmental factors, may be important determinants in the risk of liver cancer development in AFB1-exposed populations.
Kirby
G M
GM
Unit of Mechanisms of Carcinogenesis, International Agency for Research on Cancer, Lyon, France.
Wolf
C R
CR
Neal
G E
GE
Judah
D J
DJ
Henderson
C J
CJ
Srivatanakul
P
P
Wild
C P
CP
D000293:Adolescent; D000328:Adult; D016604:Aflatoxin B1 / Q000378:metabolism*; D000368:Aged; D003577:Cytochrome P-450 Enzyme System / Q000378:metabolism; D005260:Female; D005982:Glutathione Transferase / Q000378:metabolism; D006801:Humans; D007527:Isoenzymes / Q000378:metabolism; D008113:Liver Neoplasms / Q000453:epidemiology / Q000378:metabolism*; D008297:Male; D008862:Microsomes, Liver / Q000201:enzymology / Q000378:metabolism*; D008875:Middle Aged; D013785:Thailand / Q000453:epidemiology
D007527:Isoenzymes; D003577:Cytochrome P-450 Enzyme System; D016604:Aflatoxin B1; D005982:Glutathione Transferase