Polycyclic aromatic hydrocarbons (PAHs) and heterocyclic aromatic
amines (HCAs) ingested with food have repeatedly been suggested to be involved in the malignant transformation of colon epithelial cells. In order to test this hypothesis,
HCEC cells (SV40
large T antigen-immortalized human colon epithelial cells) were incubated with a racemic mixture of
benzo[c]phenanthrene dihydrodiol epoxides (
B[c]PhDE), extremely potent carcinogenic PAH metabolites in vivo, or with 2-hydroxyamino-1-methyl-6-phenylimidazo[4,5-
b]pyridine (
N-OH-PhIP), the N-hydroxylated metabolite of the most abundant HCA in cooked meat. First, it was shown that
HCEC cells express
sulfotransferase 1A1, which is needed to metabolize
N-OH-PhIP to the corresponding N-sulfonyloxy derivative, the direct precursor molecule of genotoxic nitrenium
ions. Thereafter, exponentially growing
HCEC cells were exposed five times to 0.1 microg (0.37 nmol)
B[c]PhDE/ml for 30 min or 0.72 microg (3 nmol)
N-OH-PhIP/ml for 24 h. Chemically treated
HCEC cells showed an enhanced saturation density and grew faster than the corresponding
solvent-treated cell cultures. After five treatment cycles,
HCEC(
B[c]PhDE) as well as
HCEC(
N-OH-PhIP) cells lost cell-cell contact inhibition and started piling up and forming foci in the culture flasks. Furthermore,
HCEC(
B[c]PhDE) and
HCEC(
N-OH-PhIP) cells were injected i.m. into SCID mice. Within 6 weeks after injection, eight animals out of eight injected with
HCEC(
B[c]PhDE) or
HCEC(
N-OH-PhIP) cells developed
tumors at the site of injection, thus demonstrating the high tumorigenic potential of the
HCEC(
B[c]PhDE) and
HCEC(
N-OH-PhIP) cell cultures. Taken together, we show for the first time that the abovementioned active PAH metabolites as well as
N-OH-PhIP are indeed able to malignantly transform human colon epithelial cells in vitro.