caa a22 4500 445795018 CHVBK 20180317145122.0 cr unu---uuuuu 170323e20110301xx s 000 0 eng 10.1007/s10620-010-1369-0 doi (NATIONALLICENCE)springer-10.1007/s10620-010-1369-0 Identification of GPX3 Epigenetically Silenced by CpG Methylation in Human Esophageal Squamous Cell Carcinoma [Elektronische Daten] [Yuanlong He, Yongjun Wang, Peng Li, Shengtao Zhu, Junxiong Wang, Shutian Zhang] Background: Esophageal squamous cell carcinoma (ESCC) is one of the most common causes of cancer mortality in the gastrointestinal tract. Promoter hypermethylation of tumor suppressor genes contributes to gene inactivation during development of ESCC. Aim: To identify novel methylation-silenced genes in ESCC. Methods: Genome-wide microarrays were applied to search for genes that were markedly upregulated after treatment with 5-aza-2′-deoxycytidine (5-Aza-dC) and that were markedly decreased in tumor tissue compared with paired adjacent nontumor tissue. Reverse-transcription polymerase chain reaction (PCR), immunohistochemistry, methylation-specific PCR, and bisulfite genomic sequencing were employed to investigate expression and methylation of candidate genes in five human ESCC cell lines, two human immortalized normal esophageal epithelial cell lines, primary ESCC tumor tissues, and paired adjacent nontumor tissues. Results: GPX3 was selected as a novel candidate hypermethylated gene in ESCC through microarray analysis. In most ESCC cell lines, GPX3 messenger RNA (mRNA) expression was downregulated and the CpG island of GPX3 promoter was methylated. Demethylation treatment with 5-Aza-dC restored GPX3 mRNA expression. Methylation of GPX3 promoter was more frequent in ESCC tumor tissues (71.4%) than in adjacent nontumor tissues (10.7%) (P<0.001), and methylation of GPX3 promoter correlated significantly with GPX3 mRNA downregulation. Finally, GPX3 protein expression was also significantly lower in ESCC tumor tissues than in adjacent nontumor tissues. Conclusion: GPX3 is downregulated through promoter hypermethylation in ESCC, which may be a potential biomarker of ESCC. Springer Science+Business Media, LLC, 2010 DNA methylation nationallicence Esophageal squamous cell carcinoma nationallicence GPX3 nationallicence Microarray analysis nationallicence ESCC : Esophageal squamous cell carcinoma nationallicence GPX3 : Glutathione peroxidase 3 nationallicence UCHL1 : Ubiquitin carboxyl-terminal esterase L1 nationallicence TSPYL5 : TSPY-like 5 nationallicence PPP1R14A : Protein phosphatase 1 regulatory (inhibitor) subunit 14A nationallicence DLC1 : Deleted in liver cancer 1 nationallicence 5-Aza-dC : 5-Aza-2′-deoxycytidine nationallicence ECRG4 : Esophageal cancer related gene 4 protein nationallicence APC : Adenomatous polyposis coli nationallicence HPP1 : Hyperplastic polyposis protein 1 nationallicence RT-PCR : Reverse-transcription PCR nationallicence GAPDH : Glyceraldehyde-3-phosphate dehydrogenase nationallicence MSP : Methylation-specific PCR nationallicence He Yuanlong Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut Wang Yongjun Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut Li Peng Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut Zhu Shengtao Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut Wang Junxiong Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut Zhang Shutian Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut Digestive Diseases and Sciences Springer US; http://www.springer-ny.com 56/3(2011-03-01), 681-688 0163-2116 56:3<681 2011 56 10620 https://doi.org/10.1007/s10620-010-1369-0 text/html Onlinezugriff via DOI 1 research-article jats NATIONALLICENCE 856 40 https://doi.org/10.1007/s10620-010-1369-0 text/html Onlinezugriff via DOI NATIONALLICENCE 700 1- He Yuanlong Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Wang Yongjun Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Li Peng Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhu Shengtao Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Wang Junxiong Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut NATIONALLICENCE 700 1- Zhang Shutian Department of Gastroenterology, Beijing Friendship Hospital Affiliated to the Capital Medical University, Beijing Digestive Disease Center, 95 Yong'an Road, 100050, Beijing, People's Republic of China aut NATIONALLICENCE 773 0- Digestive Diseases and Sciences Springer US; http://www.springer-ny.com 56/3(2011-03-01), 681-688 0163-2116 56:3<681 2011 56 10620 Metadata rights reserved Springer special CC-BY-NC licence nationallicence BK010053 XK010053 XK010000 NATIONALLICENCE NATIONALLICENCE NL-springer