Metabolic activation and cytogenetic effects of 2-amino-1-methyl-6-phenylimidazo[4,5-b] pyridine (PhIP) in Chinese hamster ovary cells expressing murine cytochrome P450 IA2.
We have utilized Chinese hamster ovary cell lines which stably express a murine cytochrome P450IA2 (P(3)450) cDNA to characterize more fully the mechanisms of genotoxicity of heterocyclic amines derived from cooked meats. To verify that these cell lines were capable of converting promutagens into active metabolites, we studied the microsomal metabolism and cytogenetic effects of 2-amino-1-methyl-6-phenylimidazo-[4,5-b]pridine (PhIP). Microsomal preparations derived from excision repair-deficient Chinese hamster ovary cells expressing the mouse cytochrome P(3)450 cDNA (UV5P3) converted PhIP to the genotoxic N-hydroxy-PhIP metabolite. Cytotoxic activity in UV5P3 was observed at concentrations of PhIP as low as 1 microM. Cytotoxicity of PhIP was an order of magnitude lower in a matched repair-proficient cell line (5P3R2) expressing the P(3)450 cDNA. PhIP produced a concentration-dependent increase in sister chromatid exchange (SCE) in UV5P3. N-Hydroxy-PhIP, at concentrations as low as 0.1 microM, produced an increase in SCE in both UV5P3 and in UV5 cells which lack the P(3)450 cDNA. Incubation of PhIP with UV5P3 cells increased the frequency of micronuclei (MN) in cytokinesis-blocked cells. Chromatid gaps, but not aberrations also were induced by treatment with PhIP. N-Hydroxy-PhIP produced increases in MN and chromatid gaps in both UV5 and UV5P3 cell lines; chromosomal aberrations were induced in UV5P3 cells. These results suggested that UV5P3 cells metabolize sufficient quantities of PhIP to produce cytogenetic damage and further indicated that N-hydroxylation of PhIP was requisite for mammalian genotoxicity.
Biomedical Sciences Division, Lawrence Livermore National Laboratory, Livermore, CA 94551.
M H
MH
Buonarati
J D
JD
Tucker
J L
JL
Minkler
R W
RW
Wu
L H
LH
Thompson
J S
JS
Felton
D000818:Animals; D001711:Biotransformation; D002460:Cell Line; D002869:Chromosome Aberrations; D019388:Cytochrome P-450 CYP1A2; D003577:Cytochrome P-450 Enzyme System; D015870:Gene Expression; D007093:Imidazoles / Q000737:chemistry / Q000378:metabolism / Q000633:toxicity*; D015162:Micronucleus Tests; D008861:Microsomes / Q000378:metabolism; D009153:Mutagens / Q000633:toxicity*; D010088:Oxidoreductases; D011725:Pyridines / Q000633:toxicity; D012854:Sister Chromatid Exchange
D007093:Imidazoles; D009153:Mutagens; D011725:Pyridines; C060533:2-hydroxyamino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D003577:Cytochrome P-450 Enzyme System; C049584:2-amino-1-methyl-6-phenylimidazo(4,5-b)pyridine; D010088:Oxidoreductases; D019388:Cytochrome P-450 CYP1A2