M. Ridanpaa et K. Husgafvelpursiainen, DENATURING GRADIENT GEL-ELECTROPHORESIS (DGGE) ASSAY FOR K-RAS AND N-RAS GENES - DETECTION OF K-RAS POINT MUTATIONS IN HUMAN LUNG-TUMOR DNA, Human molecular genetics, 2(6), 1993, pp. 639-644
Point mutations in the ras oncogenes are very common in lung cancers a
s well as in many of the other solid tumours. To effectively examine t
he occurrence of these mutations in a large number of tumour samples,
we have applied denaturing gradient gel electrophoresis (DGGE) for the
analysis of point mutations of the K-ras and N-ras genes, using GC-cl
amped, PCR-amplified DNA fragments. Among the 68 tumour DNA samples, w
e detected 14 mutations in the K-ras gene. This was 78% of the mutatio
ns identified by oligonucleotide hybridization. Altogether, eight of t
he nine different kinds of base substitutions found in the tumour samp
les were detected by the DGGE assay, representing substitutions at cod
ons 12, 13, and 61 of the K-ras gene. Six of the detected mutations we
re guanine to thymine transversions at codon 12; this was the most com
mon type of alteration. On the basis of our experience, the present no
n-radioactive DGGE analysis seems to be readily applicable for detecti
on of the mutations in the K-ras and N-ras genes. Types of ras gene mu
tations frequent in adenocarcinomas of the lung are also discussed.