Promoter Hypermethylation of the RUNX3 Gene in Esophageal Squamous Cell Carcinoma

Abstract

Alteration in transforming growth factor-beta (TGF-beta) signaling pathway is one of the main causes of esophageal squamous cell carcinoma (ESCC). The human runt-related transcription factor 3 (RUNX3), an important component of TGF-beta pathway which is located at 1p36, is commonly deleted in a variety of human cancers, including ESCC. Hypermethylation of RUNX3 promoter was frequently found in gastrointestinal cancers, including those of stomach, liver, colon and pancreas. However, RUNX3 promoter methylation status in ESCC has not been studied. The aim of this study was to determine whether promoter methylation of the RUNX3 gene correlates with ESCC tumor progression.Accordingly, we first determined RUNX3 mRNA expression and methylation status of its promoter region in 42 primary tumors with ESCC and Eca-109, an ESCC cell line. Loss of RUNX3 mRNA expression was detected by RT-PCR in 23 out of 42 (54.8%) ESCC specimens and Eca-109 cells. The Promoter hypermethylation was detected by Methylation Specific Polymerase Chain Reaction (MS-PCR) in 27 out of 42 (64.3%) ESCC specimen and Eca-109 cells. Importantly, we found positive correlations, not only between the promoter hypermethylation and tumor clinical pathologic stages (P = 0.003), but also between the loss of RUNX3 mRNA expression and the tumor progression (P = 0.016). Finally, we observed that the loss of RUNX3 mRNA expression is statistically correlated with the promoter hypermethylation in these tumors (P < 0.001). Our results suggest that epigenetic silencing of RUNX3 gene expression by promoter hypermethylation may play an important role in ESCC development.

Alteration of Runt-related Transcription Factor 3 Gene Expression and Biologic Behavior of Esophageal Carcinoma TE-1 Cells after 5-Azacytidine Intervention

“…West blotting assay also demonstrated that RUNX3 gene was silenced in esophageal squamous carcinoma TE-1 cells. Our results are similar to studies in hepatocellular cancer (Park et al, 2005), esophageal squamous carcinoma cells (Long et al, 2007), breast cancer (Subramaniam et al, 2009) and gastric cancer (Tang et al, 2012). But repots of Smith et al, are different.…”

“…But our experiment results are similar to those of the majority of studies (Long et al, 2007). We demonstrated that loss of RUNX3 gene expression may result from hypermethylation in TE-1 cells.…”

“…However, in Barrett's esophagus and esophageal carcinoma tissues, expression of RUNX3 gene was significantly silenced (Smith et al, 2008). Lose of RUNX3 gene expression occurs both in esophageal adenocarcinoma cells (Smith et al, 2008) and esophageal squamous carcinoma cells (Long et al, 2007). But CpG island methylator phenotype (CIMP) in esophageal carcinoma is unclear.…”

Detection of regional DNA methylation using DNA-graphene affinity interactions

“…Also, FAM134B has been reported to have roles in pathogenesis as well as prediction of prognosis of other gastrointestinal cancer, namely colorectal cancer (Kasem et al, 2014). Most current approaches detect DNA methylation in oesophageal cancers via methylation specific PCR amplification process (Kuroki et al, 2003;Long et al, 2007). In this proof-of-concept study, we have used graphene-DNA affinity interaction for detecting gene-specific DNA methylation in FAM134B.…”

Epigenetic aberrations during oncogenesis

“…Promoter hypermethylationinduced silencing of transcription factors, such as RUNX3 in esophageal cancer [32] and GATA-4 and -5 in colorectal and gastric cancer [33], leads to inactivation of their downstream targets. Silencing of DNA repair genes like BRCA1 blocks the repair of genetic mistakes and enables cells to accumulate further genetic lesions leading to neoplastic transformation of the cell [34].…”