Mutations of BRAF are associated with extensive hMLH1 promoter methylation in sporadic colorectal carcinomas

Int J Cancer. 2004 Jan 10;108(2):237-42. doi: 10.1002/ijc.11523.


Activating mutations of BRAF have been frequently observed in microsatellite unstable (MSI+) colorectal carcinomas (CRCs), in which mutations of BRAF and KRAS are mutually exclusive. Previously, we reported that hypermethylation of hMLH1 might play an important role in the tumorigenesis of right-sided sporadic CRCs with MSI showing less frequency of KRAS/TP53 alteration. Therefore, we have assumed that BRAF mutations might be highly associated with hMLH1 methylation status rather than MSI status. In this study, mutations of BRAF and KRAS and their relationship with MSI and hMLH1 methylation status were examined in 140 resected specimens of CRC. The methylation status was classified into 3 types: full methylation (FM), partial methylation (PM) and nonmethylation (NM). Only FM closely linked to reduced expression of hMLH1 protein. BRAF mutations were found in 16 cases (11%), all leading to the production of BRAF(V599E). As for MSI status, BRAF mutations were found in 43% of MSI+ and 4% of MSI- cases (p < 0.0001). Among the MSI+ individuals, BRAF mutations were more frequent in cases with hMLH1 deficiency (58%) than those with hMSH2 deficiency (0%; p=0.02). Moreover, they were found in 69% of FM, 4% of PM and 4% of NM, revealing a striking difference between FM and the other 2 groups (FM vs. PM or NM; p < 0.0001). These findings suggest that BRAF activation may participate in the carcinogenesis of sporadic CRCs with hMLH1 hypermethylation in the proximal colon, independently of KRAS activation.

MeSH terms

  • Adaptor Proteins, Signal Transducing
  • Adenocarcinoma / genetics
  • Adenocarcinoma, Mucinous / genetics
  • Adult
  • Aged
  • Aged, 80 and over
  • Base Pair Mismatch
  • Carrier Proteins
  • Colorectal Neoplasms / genetics*
  • Cytoskeletal Proteins / genetics
  • DNA Methylation*
  • DNA, Neoplasm / genetics
  • DNA-Binding Proteins*
  • Female
  • Gene Expression Regulation, Neoplastic
  • Genes, ras / physiology
  • Humans
  • Male
  • Microsatellite Repeats
  • Middle Aged
  • MutL Protein Homolog 1
  • MutS Homolog 2 Protein
  • Mutation / genetics*
  • Neoplasm Proteins / genetics*
  • Neoplasm Staging
  • Nuclear Proteins
  • Oncogene Proteins / genetics*
  • Promoter Regions, Genetic / genetics*
  • Proto-Oncogene Proteins / genetics*
  • Proto-Oncogene Proteins B-raf
  • Trans-Activators / genetics
  • beta Catenin


  • Adaptor Proteins, Signal Transducing
  • CTNNB1 protein, human
  • Carrier Proteins
  • Cytoskeletal Proteins
  • DNA, Neoplasm
  • DNA-Binding Proteins
  • MLH1 protein, human
  • Neoplasm Proteins
  • Nuclear Proteins
  • Oncogene Proteins
  • Proto-Oncogene Proteins
  • Trans-Activators
  • beta Catenin
  • BRAF protein, human
  • Proto-Oncogene Proteins B-raf
  • MSH2 protein, human
  • MutL Protein Homolog 1
  • MutS Homolog 2 Protein