Gene Promoter and Exon DNA Methylation Changes in Colon Cancer Development - MRNA Expression and Tumor Mutation Alterations

Overview

Journal BMC Cancer

Publisher Biomed Central

Specialty Oncology

Date 2018 Jun 28

PMID 29945573

Citations 32

Authors

Bela Molnar

Orsolya Galamb

Balint Peterfia

Barnabas Wichmann

Istvan Csabai

Andras Bodor

Alexandra Kalmar

Krisztina Andrea Szigeti

Barbara Kinga Bartak

Zsofia Brigitta Nagy

Gabor Valcz

Arpad V Patai

Peter Igaz

Zsolt Tulassay

Affiliations

Soon will be listed here.

Abstract

Background: DNA mutations occur randomly and sporadically in growth-related genes, mostly on cytosines. Demethylation of cytosines may lead to genetic instability through spontaneous deamination. Aims were whole genome methylation and targeted mutation analysis of colorectal cancer (CRC)-related genes and mRNA expression analysis of TP53 pathway genes.

Methods: Long interspersed nuclear element-1 (LINE-1) BS-PCR followed by pyrosequencing was performed for the estimation of global DNA metlyation levels along the colorectal normal-adenoma-carcinoma sequence. Methyl capture sequencing was done on 6 normal adjacent (NAT), 15 adenomatous (AD) and 9 CRC tissues. Overall quantitative methylation analysis, selection of top hyper/hypomethylated genes, methylation analysis on mutation regions and TP53 pathway gene promoters were performed. Mutations of 12 CRC-related genes (APC, BRAF, CTNNB1, EGFR, FBXW7, KRAS, NRAS, MSH6, PIK3CA, SMAD2, SMAD4, TP53) were evaluated. mRNA expression of TP53 pathway genes was also analyzed.

Results: According to the LINE-1 methylation results, overall hypomethylation was observed along the normal-adenoma-carcinoma sequence. Within top50 differential methylated regions (DMRs), in AD-N comparison TP73, NGFR, PDGFRA genes were hypermethylated, FMN1, SLC16A7 genes were hypomethylated. In CRC-N comparison DKK2, SDC2, SOX1 genes showed hypermethylation, while ERBB4, CREB5, CNTN1 genes were hypomethylated. In certain mutation hot spot regions significant DNA methylation alterations were detected. The TP53 gene body was addressed by hypermethylation in adenomas. APC, TP53 and KRAS mutations were found in 30, 15, 21% of adenomas, and in 29, 53, 29% of CRCs, respectively. mRNA expression changes were observed in several TP53 pathway genes showing promoter methylation alterations.

Conclusions: DNA methylation with consecutive phenotypic effect can be observed in a high number of promoter and gene body regions through CRC development.

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