Promoter hypermethylation of CDH13 is a common, early event in human esophageal adenocarcinogenesis and correlates with clinical risk factors

Abstract

Although the CDH13 gene has been shown to undergo epigenetic silencing by promoter methylation in many types of tumors, hypermethylation of this gene in Barrett's-associated esophageal adenocarcinogenesis has not been studied. Two hundred fifty-nine human esophageal tissues were therefore examined for CDH13 promoter hypermethylation by real-time methylation-specific PCR. CDH13 hypermethylation showed discriminative receiveroperator characteristic curve profiles, sharply demarcating esophageal adenocarcinoma (EAC) from esophageal squamous cell carcinoma (ESCC) and normal esophagus (NE) (p < 0.0001). CDH13 normalized methylation values (NMV) were significantly higher in Barrett's esophagus (BE), dysplastic BE (D) and EAC than in NE (p < 0.0000001). CDH13 hypermethylation frequency was 0% in NE but increased early during neoplastic progression, rising to 70% in BE, 77.5% in D and 76.1% in EAC. Both CDH13 hypermethylation frequency and its mean NMV were significantly higher in BE with than without accompanying EAC. In contrast, only 5 (19.2%) of 26 ESCCs exhibited CDH13 hypermethylation. Furthermore, both CDH13 hypermethylation frequency and its mean NMV were significantly higher in EAC than in ESCC, as well as in BE or D vs. ESCC. Interestingly, mean CDH13 NMV was significantly lower in short-segment than in long-segment BE, a known clinical risk factor for neoplastic progression. Similarly, BE segment length was significantly lower in specimens with unmethylated than with methylated CDH13 promoters. 5-aza-2 0 -deoxycytidine treatment of OE33 EAC and KYSE220 ESCC cells reduced CDH13 methylation and increased CDH13 mRNA expression. These findings suggest that hypermethylation of CDH13 is a common, tissue-specific event in human EAC, occurs early during BE-associated neoplastic progression, and correlates with known clinical neoplastic progression risk factors. ' 2008 Wiley-Liss, Inc.Key words: CDH13; hypermethylation; EAC; ESCC CDH13 (also known as H-cadherin and T-cadherin), a member of the cadherin gene superfamily, was isolated and has been mapped to 16q24, 1 a locus that frequently undergoes deletion in human cancers, including esophageal carcinoma. 2,3 In contrast to other known cadherins such as E-cadherin, N-cadherin and P-cadherin, which are transmembrane proteins, CDH13 lacks conventional transmembrane and cytoplasmic domains and is attached to the plasma membrane through a glycosyl phosphatidyl inositol anchor.1,4-6 Several studies have suggested that CDH13 functions as a tumor suppressor gene and possesses potent antitumor activity in several human cancers both in vitro and in vivo.7-10 Overexpression of CDH13 in human breast carcinoma cells (MDAMB435) reduced their invasive potential in vitro and tumor formation in vivo, accompanied by reversion from invasive to normal cell morphology.7 Loss of CDH13 protein expression is associated with tumorigenicity of human non-small cell lung cancer cells. 8,9 In cutaneous squamous cell carcinoma cells, overexpression of CDH13 induced a delay in the G 2 /M phase of...

Involvement of Members of the Cadherin Superfamily in Cancer

“…Highly invasive growth pattern (Berx et al 1998) Up-regulated expression Epithelial ovarian cancer (Sato et al 1998;Maruyama et al 2001;Toyooka et al 2001;Toyooka et al 2002;Roman-Gomez et al 2003;Hibi et al 2004;Sakai et al 2004;Kim et al 2005;Wang et al 2007;Brock et al 2008;Jin et al 2008;Yan et al 2008) LOH and promoter methylation Ovary Skin (BCC and SCC) Diffuse large B cell lymphoma HCC Tumor invasion; increased cell proliferation and resistance to apoptosis (Kawakami et al 1999;Takeuchi et al 2002a;Takeuchi et al 2002b;Ogama et al 2004;Chan et al 2008) Expression in intratumoral endothelium…”

Hypermethylation of collagen α2 (I) gene (COL1A2) is an independent predictor of survival in head and neck cancer

“…The PCR products were 82 bp long. To analyze the methylation status of the p16 gene [5], E-cadherin gene [6], RASSF1A gene [7], H-cadherin gene [22], DAPK gene [23], MGMT gene [23], and DCC gene [24], primers and conditions as described previously were used. The PCR products were separated by electrophoresis through a 9% polyacrylamide gel and stained with ethidium bromide.…”

Chemoprevention in Barrett’s Esophagus

“…Contrary to this, epigenetic effects, such as global hypomethylation and promoterspecific hypermethylation that are more dynamic in nature, or microRNA will likely be better markers to assess the efficacy of a chemopreventive agent. 58,[104][105][106][107][108][109] Recent studies have demonstrated progressive increase in the levels of GLI-1, vascular endothelial growth factor, transcription factor MCM2, COX-2 enzyme, PGE2, polyamine, 3,87 and upregulation of prosurvival NF-kB during oncogenesis in BE. 108 These biochemical and molecular markers of neoplastic progression also can assess if the molecular pathways intended to be targeted by chemopreventive agents are actually being downregulated.…”