The role of EZH2 and DNA methylation in the silencing of the tumour suppressor RUNX3 in colorectal cancer.

Carcinogenesis
Liudmila L KodachJames C H Hardwick

Abstract

In gastric cancer, a new epigenetic mechanism of tumour suppressor loss has been suggested where the histone methyltransferase enhancer of zeste homolog 2 (EZH2) is responsible for loss of expression of RUNX3. This is consistent with EZH2 upregulation in multiple cancer types being associated with poor prognosis. We investigated whether EZH2 influences the expression of RUNX3 in colorectal cancer (CRC) and whether this is independent of methylation. We determined protein and messenger RNA (mRNA) levels of EZH2 and RUNX3 and assessed RUNX3 methylation with methylation-specific polymerase chain reaction using 72 human CRCs and 8 CRC cell lines. We assessed the effect of efficient RNA interference-mediated knockdown of EZH2 on RUNX3 levels, cell viability and H3K27 trimethylation of the RUNX3 promoter using chromatin immunoprecipitation. Despite higher levels of EZH2 and lower levels of RUNX3 in CRC specimens in general, no inverse correlation between EZH2 and RUNX3 in paired samples was found arguing against a major role for histone methylation in silencing RUNX3 in CRC. Conversely, downregulation of RUNX3 mRNA in the same tumours was associated with RUNX3 DNA methylation (P < 0.05). In cell lines, knockdown of EZH2 removed the r...Continue Reading

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Citations

Jul 19, 2011·PloS One·Bettina FussbroichFelix Hoppe-Seyler
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Methods Mentioned

BETA
acetylation
protein assay
electrophoresis
PCR
transfection
immunoprecipitation
PCRs
immunoprecipitations

Software Mentioned

Statistical Package of Social Science ( SPSS )

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