The first study to address DAPT methylation changes in ESCC in a large set of genes, but the microarray they used is the Illumina GoldenGate Methylation assay which only covers 1505 CpG sites. Infinium Methylation 450K array is a newly developed BeadChip platform, which can test more than 480,000 individual CpG sites in the human genome. The high correlation between Infinium Methylation 450K array data and whole-genome bisulfate sequencing data indicates that this new BeadChip can provide reliable DNA methylation data for epigenomic profiling studies. Furthermore, aberrant DNA methylation usually occurs somatically in cancers and can be detected in the blood circulation, so it may serve as a novel marker for cancer. For example, the methylation of some tumor related genes such as p16, DAPK, RAR-b, CDH1, and RASSF1A have been detected in circulating cell-free DNA. In the current study, we analyzed global methylation profiling of ESCC and normal adjacent tissue in Chinese cancer patients and esophageal mucosa from Chinese healthy individuals, using Infinium Methylation 450K array. After analysis of the methylation differences and then in combination with independent gene expression data using BRB -Array Tools of the National Cancer Institute, a set of genes that are deregulated by aberrant DNA methylation in ESCC was identified. We then focused on 3 aberrant DNA methylation genes—EPB41L3, GPX3, and COL14A1, with validation analysis using additional ESCC tumor tissues and adjacent normal tissues. In order to evaluate whether the methylation status of the 3 candidate genes is useful for diagnosing ESCC, we also tested the methylation of the circulating cell-free DNA in patients’ plasma and controls. All participants signed an informed consent and information was obtained using a standardized questionnaire, including data on tobacco smoking, alcohol drinking, and family history. An individual was classified as a smoker if he or she smoked at least one cigarette per day for more than one year. An individual was classified as a drinker if he or she drank alcoholic beverages five times per week for more than one year. Family history of cancer was defined as positive when at least one of the patient’s firstdegree relatives was definitely diagnosed with cancer. This study was approved by the ethics committee of the Anyang Cancer Hospital in Henan province in China. Written Informed consent have been obtained from all the participants and kept as records submitted back to ethics committee. For minor participants less than 18 years old, written informed consent was obtained from guardians. The ethics committee approved the consent procedure. Although epigenetic events have been implicated in esophageal cancer, genome-wide epigenetic deregulation and precise targets of aberrant DNA methylation during the development and progression of ESCC have not been defined. In this pilot study, we describe the methylation profile of Chinese ESCC patients. One hundred and sixty eight differentially methylated genes that were functionally deregulated in ESCC were identified.