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...