O Zulhabri1, J Rahman, S Ismail, M R Isa, W N Wan Zurinah. 1. UKM Medical Molecular Biology Institute, Universiti Kebangsaan Malaysia Medical Centre, Jalan Yaacob Latiff, Kuala Lumpur, Malaysia. zulhabri_othman@yahoo.com.
Abstract
INTRODUCTION: K-ras gene mutations in codons 12 and 13 are one of the earliest events in colon carcinogenesis. METHODS: DNA was extracted from 25 mg of tumour tissue (n = 70) that were taken from tumour mass and pairs with normal epithelial tissue distant from the tumour of colorectal cancer patients. Exon 1 and exon 2 of the K-ras gene were amplified. Hotspot mutations were detected using polymerase chain reaction-based single-strand conformation polymorphism method and confirmed by direct DNA sequencing analysis. RESULTS: Mutations were identified in 14 out of the 70 (20%) colorectal carcinoma tissues. Single-base transition from GGT to GAT (glycine to aspartate) in codon 12 was detected in nine samples, while three samples presented with GGC to GAC transition in codon 13. Patients with large adenoma had a 12-fold higher likelihood of K-ras mutations (odds ratios [OR] 12.31; 95% confidence intervals [CI] 1.81-83.76). Tumours located at the left colon were more likely to present with K-ras mutations (OR 4.54; 95% CI 0.96-21.54). CONCLUSION: Our study showed a high frequency of G to A transition of codon 12 mutation of the K-ras gene, with significant correlation with tumour size and tumour location.
INTRODUCTION:K-ras gene mutations in codons 12 and 13 are one of the earliest events in colon carcinogenesis. METHODS: DNA was extracted from 25 mg of tumour tissue (n = 70) that were taken from tumour mass and pairs with normal epithelial tissue distant from the tumour of colorectal cancerpatients. Exon 1 and exon 2 of the K-ras gene were amplified. Hotspot mutations were detected using polymerase chain reaction-based single-strand conformation polymorphism method and confirmed by direct DNA sequencing analysis. RESULTS: Mutations were identified in 14 out of the 70 (20%) colorectal carcinoma tissues. Single-base transition from GGT to GAT (glycine to aspartate) in codon 12 was detected in nine samples, while three samples presented with GGC to GAC transition in codon 13. Patients with large adenoma had a 12-fold higher likelihood of K-ras mutations (odds ratios [OR] 12.31; 95% confidence intervals [CI] 1.81-83.76). Tumours located at the left colon were more likely to present with K-ras mutations (OR 4.54; 95% CI 0.96-21.54). CONCLUSION: Our study showed a high frequency of G to A transition of codon 12 mutation of the K-ras gene, with significant correlation with tumour size and tumour location.
Authors: Joanna H M Tong; Raymond W M Lung; Frankie M C Sin; Peggy P Y Law; Wei Kang; Anthony W H Chan; Brigette B Y Ma; Tony W C Mak; Simon S M Ng; Ka Fai To Journal: Cancer Biol Ther Date: 2014-03-18 Impact factor: 4.742