Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.

A CpG island methylator phenotype (CIMP) is displayed by a distinct subset of colorectal cancers with a high frequency of DNA hypermethylation in a specific group of CpG islands. Recent studies have shown that an activating mutation of BRAF (BRAF(V600E)) is tightly associated with CIMP, raising the...

Full description

Saved in:
Bibliographic Details
Main Authors: Toshinori Hinoue, Daniel J Weisenberger, Fei Pan, Mihaela Campan, Myungjin Kim, Joanne Young, Vicki L Whitehall, Barbara A Leggett, Peter W Laird
Format: Article
Language:English
Published: Public Library of Science (PLoS) 2009-12-01
Series:PLoS ONE
Online Access:https://europepmc.org/articles/pmc2791229?pdf=render
Tags: Add Tag
No Tags, Be the first to tag this record!
_version_ 1850133889772683264
author Toshinori Hinoue
Daniel J Weisenberger
Fei Pan
Mihaela Campan
Myungjin Kim
Joanne Young
Vicki L Whitehall
Barbara A Leggett
Peter W Laird
author_facet Toshinori Hinoue
Daniel J Weisenberger
Fei Pan
Mihaela Campan
Myungjin Kim
Joanne Young
Vicki L Whitehall
Barbara A Leggett
Peter W Laird
author_sort Toshinori Hinoue
collection DOAJ
description A CpG island methylator phenotype (CIMP) is displayed by a distinct subset of colorectal cancers with a high frequency of DNA hypermethylation in a specific group of CpG islands. Recent studies have shown that an activating mutation of BRAF (BRAF(V600E)) is tightly associated with CIMP, raising the question of whether BRAF(V600E) plays a causal role in the development of CIMP or whether CIMP provides a favorable environment for the acquisition of BRAF(V600E). We employed Illumina GoldenGate DNA methylation technology, which interrogates 1,505 CpG sites in 807 different genes, to further study this association. We first examined whether expression of BRAF(V600E) causes DNA hypermethylation by stably expressing BRAF(V600E) in the CIMP-negative, BRAF wild-type COLO 320DM colorectal cancer cell line. We determined 100 CIMP-associated CpG sites and examined changes in DNA methylation in eight stably transfected clones over multiple passages. We found that BRAF(V600E) is not sufficient to induce CIMP in our system. Secondly, considering the alternative possibility, we identified genes whose DNA hypermethylation was closely linked to BRAF(V600E) and CIMP in 235 primary colorectal tumors. Interestingly, genes that showed the most significant link include those that mediate various signaling pathways implicated in colorectal tumorigenesis, such as BMP3 and BMP6 (BMP signaling), EPHA3, KIT, and FLT1 (receptor tyrosine kinases) and SMO (Hedgehog signaling). Furthermore, we identified CIMP-dependent DNA hypermethylation of IGFBP7, which has been shown to mediate BRAF(V600E)-induced cellular senescence and apoptosis. Promoter DNA hypermethylation of IGFBP7 was associated with silencing of the gene. CIMP-specific inactivation of BRAF(V600E)-induced senescence and apoptosis pathways by IGFBP7 DNA hypermethylation might create a favorable context for the acquisition of BRAF(V600E) in CIMP+ colorectal cancer. Our data will be useful for future investigations toward understanding CIMP in colorectal cancer and gaining insights into the role of aberrant DNA hypermethylation in colorectal tumorigenesis.
format Article
id doaj-art-03642a8598fe4608b084e418e3417e66
institution OA Journals
issn 1932-6203
language English
publishDate 2009-12-01
publisher Public Library of Science (PLoS)
record_format Article
series PLoS ONE
spelling doaj-art-03642a8598fe4608b084e418e3417e662025-08-20T02:31:51ZengPublic Library of Science (PLoS)PLoS ONE1932-62032009-12-01412e835710.1371/journal.pone.0008357Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.Toshinori HinoueDaniel J WeisenbergerFei PanMihaela CampanMyungjin KimJoanne YoungVicki L WhitehallBarbara A LeggettPeter W LairdA CpG island methylator phenotype (CIMP) is displayed by a distinct subset of colorectal cancers with a high frequency of DNA hypermethylation in a specific group of CpG islands. Recent studies have shown that an activating mutation of BRAF (BRAF(V600E)) is tightly associated with CIMP, raising the question of whether BRAF(V600E) plays a causal role in the development of CIMP or whether CIMP provides a favorable environment for the acquisition of BRAF(V600E). We employed Illumina GoldenGate DNA methylation technology, which interrogates 1,505 CpG sites in 807 different genes, to further study this association. We first examined whether expression of BRAF(V600E) causes DNA hypermethylation by stably expressing BRAF(V600E) in the CIMP-negative, BRAF wild-type COLO 320DM colorectal cancer cell line. We determined 100 CIMP-associated CpG sites and examined changes in DNA methylation in eight stably transfected clones over multiple passages. We found that BRAF(V600E) is not sufficient to induce CIMP in our system. Secondly, considering the alternative possibility, we identified genes whose DNA hypermethylation was closely linked to BRAF(V600E) and CIMP in 235 primary colorectal tumors. Interestingly, genes that showed the most significant link include those that mediate various signaling pathways implicated in colorectal tumorigenesis, such as BMP3 and BMP6 (BMP signaling), EPHA3, KIT, and FLT1 (receptor tyrosine kinases) and SMO (Hedgehog signaling). Furthermore, we identified CIMP-dependent DNA hypermethylation of IGFBP7, which has been shown to mediate BRAF(V600E)-induced cellular senescence and apoptosis. Promoter DNA hypermethylation of IGFBP7 was associated with silencing of the gene. CIMP-specific inactivation of BRAF(V600E)-induced senescence and apoptosis pathways by IGFBP7 DNA hypermethylation might create a favorable context for the acquisition of BRAF(V600E) in CIMP+ colorectal cancer. Our data will be useful for future investigations toward understanding CIMP in colorectal cancer and gaining insights into the role of aberrant DNA hypermethylation in colorectal tumorigenesis.https://europepmc.org/articles/pmc2791229?pdf=render
spellingShingle Toshinori Hinoue
Daniel J Weisenberger
Fei Pan
Mihaela Campan
Myungjin Kim
Joanne Young
Vicki L Whitehall
Barbara A Leggett
Peter W Laird
Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.
PLoS ONE
title Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.
title_full Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.
title_fullStr Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.
title_full_unstemmed Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.
title_short Analysis of the association between CIMP and BRAF in colorectal cancer by DNA methylation profiling.
title_sort analysis of the association between cimp and braf in colorectal cancer by dna methylation profiling
url https://europepmc.org/articles/pmc2791229?pdf=render
work_keys_str_mv AT toshinorihinoue analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT danieljweisenberger analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT feipan analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT mihaelacampan analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT myungjinkim analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT joanneyoung analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT vickilwhitehall analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT barbaraaleggett analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling
AT peterwlaird analysisoftheassociationbetweencimpandbrafincolorectalcancerbydnamethylationprofiling