Specific PCR Amplification for N-ras Mutations in Neoplastic Thyroid Diseases

Abstract

We analyzed point mutations of N-ras protooncogene codon 61 in thyroid neoplasms by means of a mutation-specific PCR method. In this method, one of the paired primers has a base at the 3' terminal that is complementary to a mutated base of the DNA sequence to be analyzed. With this primer, only alleles which have the same mutation can be amplified. Among 24 thyroid tissues, we detected 2 point mutations out of 7 follicular carcinomas (29%). One tumor had a cytosine to adenine substitution mutation at the first base of codon 61, and the other had an adenine to guanine substitution mutation in the second base of the same codon. The same mutations were not detected in 7 follicular adenomas or 1 papillary carcinoma. These results were confirmed by both dot blot hybridization and direct sequencing method. Mutation of N-ras codon 61 may be significant in malignant transformation of follicular thyroid tumors. Because of its easy availability, the mutation-specific PCR method is a useful screening test for N-ras mutations.