Aberrant promoter methylation of theTPEFgene in esophageal squamous cell carcinoma

Abstract

Aberrant methylation of tumor suppressor genes plays an important role in the development of esophageal squamous cell carcinoma (ESCC). The purpose of the present study was to identify the epigenetic changes in ESCC. Methylation-sensitive arbitrarily primed polymerase chain reaction (MS AP-PCR) analysis was used on 22 matched ESCC tumors and adjacent normal tissues. Through this screen we identified a frequently methylated fragment that showed a high homology to the 5'-CpG island of the gene encoding a transmembrane protein containing epidermal growth factor and follistatin domains (TPEF). The methylation status of the TPEF gene was then detected by bisulfite sequencing and the levels of TPEF mRNA were detected by RT-PCR. In addition, the effects of a methylation inhibitor 5-aza-2'-deoxycytidine on TPEF mRNA expression was determined in cells of ESCC cell lines. Hypermethylation of the 5'-CpG island of TPEF was found in 12 of 22 (54.5%) primary tumors. Reverse transcription PCR analysis demonstrated that TPEF mRNA expression was significantly lower in tumors than in adjacent normal tissues, which is associated with promoter hypermethylation. In addition, treatment of ESCC cell lines with 5-aza-2'-deoxycytidine led to re-expression of the TPEF transcript. In conclusion, we observed promoter of TPEF gene is frequently hpermethylated, and is associated with the loss of TPEF mRNA expression in ESCC samples. Promoter hypermethylation of TPEF gene may play a role in the development of ESCC.

Identification of GPX3 Epigenetically Silenced by CpG Methylation in Human Esophageal Squamous Cell Carcinoma

“…Epigenetic changes such as promoter DNA methylation can induce inactivation of tumor suppressor genes that are important to prevent carcinogenesis. Recently, a number of genes, e.g., ECRG4 [4], p16 [5], HPP1 [6], DLC1 [7], UCHL1 [8], and TSPYL5 and PPP1R14A [9], have been found to be aberrantly methylated in ESCC. Nevertheless, novel and specific methylation-sensitive tumor suppressor genes in ESCC remain to be identified.…”

Nutlin‐3 acts as a DNA methyltransferase inhibitor to sensitize esophageal cancer to chemoradiation

“…We next verify whether repression of DNMT caused by Nutlin‐3 could decrease DNA methylation level in ESCC cells. We examined the DNA methylation levels of 14 TSGs that were reported to be associated with poor ESCC prognosis when hypermethylated, including FHIT, SOX17, SLIT2, RARβ, PAX6, CHFR, MGMT, MLH1, DCC, APC, TMEFF2, SFRP1, DAPK and p16 (Supporting Information: Table ) 25–36 . Various ESCC cell lines including chemoradiation‐resistant ones (KYSE510‐R and CE81T‐R) were examined upon Nutlin‐3 treatment.…”

“…We examined the DNA methylation levels of 14 TSGs that were reported to be associated with poor ESCC prognosis when hypermethylated, including FHIT, SOX17, SLIT2, RARβ, PAX6, CHFR, MGMT, MLH1, DCC, APC, TMEFF2, SFRP1, DAPK and p16 (Supporting Information: Table S3). [25][26][27][28][29][30][31][32][33][34][35][36] Various ESCC cell lines including chemoradiationresistant ones (KYSE510-R and CE81T-R) were examined upon Nutlin-3 treatment. 5-aza-dC was included for comparison.…”

A review of the alterations in DNA methylation in esophageal squamous cell carcinoma

“…As a result, there has been considerable research carried out on this epigenetic alteration of various genes ( Table 2). Aberrant hypermethylation has been reported for the following genes: PTX3 (a member of the pentraxin superfamily) [74], SFRP1 (a Wnt signaling modulator) [75], SOCS1 (a negative regulator of the JAK/STAT pathway) [76], HSPB2 (the heat shock protein B2) [77], LTBP2 (a component of the extracellular matrix microfibrils) [78], CLDN4 (an adhesion molecule constituting tight junctions) [79], PCDH17 (a member of the protocadherin superfamily) [80], SCGB3A1 (a putative cytokine gene) [81], TMEFF2 (a transmembrane protein) [82], TIMP3 (Tissue inhibitor of metalloproteinase-3) [83,84], ENG (a homodimeric transmembrane glycoprotein) [85], UCHL1 (a neuron-specific protein; PGP9.5) [86,87], RUNX3 (a family of transcriptional factors) [88][89][90], DCC (a netrin receptor; deleted in colorectal carcinoma) [91], HOPX (a homeobox only protein) [92], GRIN1 (the first class of glutamate receptors) [93], SST (the primary inhibitor of gastrin-stimulated gastric acid secretion) [94], TAC1 (neuropeptides substance P, neurokinin A, and neuropeptide K and c) [95], EP300 (a transcriptional coactivator; p300) [96], CDX2 (the mammalian homologue of the homeobox gene) [97,98], CRABP1 (cellular retinoic acid binding protein 1) [99], MT3 (metallothionein 3) [100], RBP1 (cellular retinol-binding protein 1; retinoic acid signaling) [101], RARRES1 (tazarotene-induced gene 1; retinoic acid signaling) [101], LRP1B (low-density lipoprotein receptor-related protein 1B) [102], CADM1 (a transmembrane glycoprotein) [103] and CHFR (checkpoint with FHA and ring finger) [104].…”

“…Member of the cadherin superfamily [80] SCGB3A1 Putative cytokine gene [81] TMEFF2 Transmembrane protein [82] TIMP3 Systemic cytokine regulation [83,84] ENG Homodimeric transmembrane glycoprotein [85] UCHL1…”