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

Chin Med J (Engl). 2007 Dec 5;120(23):2119-23.

Abstract

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 1a 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 1b 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 polymorphysim.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Cell Line, Tumor
  • Checkpoint Kinase 2
  • Chromatography, High Pressure Liquid
  • Colorectal Neoplasms / genetics*
  • DNA Damage
  • Humans
  • Mutation*
  • Protein Serine-Threonine Kinases / genetics*

Substances

  • Checkpoint Kinase 2
  • CHEK2 protein, human
  • Protein Serine-Threonine Kinases