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Genetic alterations in rat colon tumors induced by heterocyclic amines

✍ Scribed by Minoru Toyota; Toshikazu Ushijima; Hideki Kakiuchi; Federico Canzian; Masatoshi Watanabe; Kohzoh Imai; Takashi Sugimura; Minako Nagao

Publisher
John Wiley and Sons
Year
1996
Tongue
English
Weight
417 KB
Volume
77
Category
Article
ISSN
0008-543X

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✦ Synopsis

BACKGROUND.

In rat colon tumors induced by the cooked food mutagens 2-aminol-methyl-6-phenylimidazo[4,5-b]pyridine (PhIP) and 2-amino-3-methylimidazo[4,5 -Qquinoline (IQ), ras and p53 alterations are rarely detected. To investigate the roles of the APC gene and microsatellite instability (MI) in PhIP-induced colon carcinogenesis, mutations of the APC gene and alterations of microsatellites were examined.

METHODS.

Complementary DNA sequence of the rat APC gene were determined by polymerase chain reaction (PCR) using primers based on the human APC sequence. PCR-single strand conformation polymorphism (SSCP) analysis was performed using primers based on sequences of flanking introns and exon 15. Microsatellite alterations were also analyzed using 85 microsatellite sequences dispersed through most of the rat chromosomes.

RESULTS.

Five mutations in the APC gene were detected in four of eight PhIPinduced rat colon tumors. All five mutations involved deletion of a guanine base in a 5'-GGGA-3' sequence. Only 2 of 13 IQ-induced colon tumors had mutations of the APC gene and these were base substitution mutations. Seven of eight PhIPinduced colon tumors had microsatellite alterations in at least one locus, whereas no alterations were observed in the IQ-induced colon tumors.

CONCLUSIONS.

The specific 5'-GGGA-3' to 5'-GGA-3' mutation and MI demonstrated in this study are strong evidence of a mutational fingerprint of PhlP.

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