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Mutation analysis of the checkpoint kinase 2 gene in colorectal cancer cell lines

Chinese Medical Journal 2007年第23期120卷 2119-2123页

作者： LIU Wei-dong ZHONG Bai-yun ZHANG Yang-de CHOI Gyu-seog National Hepatobiliary and Enteric Surgery Research Center Ministry of Health Xiangya Hospital Central-south University Changsha 410008 China Division of colorectal Surgery Department of Surgery Bio-medical Research Institute Kyungpook National University Hospital Daegu 700412 Korea

Background Checkpoint kinase 2 （CHK2） is a DNA damage-activated protein kinase which is involved in cell cycle checkpoint control, CHK2 gene could be a candidate gene for colorectal cancer susceptibility, But there are few systematic reports on mutation of CHK2 in colorectal cancer. Methods The mutations of all 14 exons of CHK2 in 56 colorectal cancer cell lines were screened systematically, using denaturing high-performance liquid chromatography （DHPLC） to screen the mismatches of the CHK2 exons amplified products, and then the suspected mutant cell lines were scanned by nucleotide sequence analysis. Results VACO400 in CHK2 exon la was suspected to have mutation by DHPLC and confirmed by sequence, but this was nonsense mutation. C106, CX-1, HT-29, SK01, SW480, SW620 and VACO400 in CHK2 exon lb were confirmed to have the same nonsense mutation in 11609 A〉G. DLD-1 and HCT-15 in CHK2 exon 2 were confirmed to have missense mutation R145W, which was heterozygous C〉T missense mutation at nucleotide 433, leading to an Arg〉Trp substitution within the FHA domain. Conclusions The CHK2 mutation in colorectal cancer is a low frequency event, There are just 10 cell lines to have sequence variations in all the 14 exons in 56 colorectal cancer cell lines and only DLD-1/HCT-15 had heterozygous missense mutation. These findings may give useful information of susceptibility of colorectal cancer as single nucleotide polvmorphvsim.

关键词： checkpoint kinase 2 colorectal cancer denaturing high-performance liquid chromatography cell line, colorectal cancer

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