title: Epigenetic inactivation of SPINT2 is associated with tumor suppressive function in esophageal squamous cell carcinoma.
abstract: Hepatocyte growth factor activator inhibitor type 2 (SPINT2), a Kunitz-type serine proteinase inhibitor, has been identified as a putative tumor suppressor gene silenced by promoter methylation. We aimed to investigate whether SPINT2 might act as an esophageal squamous cell carcinoma (ESCC) tumor suppressor gene. Four ESCC cell lines, Fifty-two ESCC tissues and twenty-nine neighboring non-cancerous tissues were included in this study. The expression of SPINT2 was monitored by real time PCR. Bisulfite genomic sequencing and methylation-specific PCR were used to analyze methylation status. The effect of SPINT2 on cell proliferation and apoptosis in EC109 and EC9706 cells was observed by CCK-8 assay and flow cytometric analysis. We found that silencing of SPINT2 was associated with promoter methylation in ESCC cell lines. The densely methylated SPINT2 promoter region was confirmed by bisulfite genomic sequencing. Ectopic expression of SPINT2 inhibited cell proliferation through inducing cell apoptosis in vitro. Furthermore, methylation-specific PCR analysis revealed that SPINT2 promoter methylation was prominent in carcinoma tissues (52.08%) compared with neighboring non-cancerous tissues (22.58%). Kaplan-Meier analysis showed that patients with SPINT2 hypermethylation had shorter survival time. The tumor suppressor gene of SPINT2 is commonly silenced by promoter hypermethylation in human ESCC and SPINT2 hypermethylation is correlated with poor overall survival, implicating SPINT2 is an underlying prognostic marker for human ESCC.
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Dongli
authors.initials: D
authors.lastname: Yue
authors.affiliation: The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Qingxia
authors.initials: Q
authors.lastname: Fan
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Xinfeng
authors.initials: X
authors.lastname: Chen
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Feng
authors.initials: F
authors.lastname: Li
authors.affiliation: The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Liping
authors.initials: L
authors.lastname: Wang
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Lan
authors.initials: L
authors.lastname: Huang
authors.affiliation: The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Wenjie
authors.initials: W
authors.lastname: Dong
authors.affiliation: The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Xiaoqi
authors.initials: X
authors.lastname: Chen
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Zhen
authors.initials: Z
authors.lastname: Zhang
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The School of Life Sciences, Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Jinyan
authors.initials: J
authors.lastname: Liu
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The School of Life Sciences, Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Fei
authors.initials: F
authors.lastname: Wang
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The Department of Gastroenterology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China.
authors.forename: Meng
authors.initials: M
authors.lastname: Wang
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The Department of Hematology/Oncology, School of Medicine, Northwestern University, Chicago 60611, USA.
authors.forename: Bin
authors.initials: B
authors.lastname: Zhang
authors.affiliation: The Biotherapy Center, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The Department of Oncology, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China; The School of Life Sciences, Zhengzhou University, Zhengzhou 450052, Henan, China; The institute of Clinical-Medicine, the First Affiliated Hospital of Zhengzhou University, Zhengzhou 450052, Henan, China. Electronic address: chyizhang@gmail.com.
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
mesh_terms: D000328:Adult; D000368:Aged; D000369:Aged, 80 and over; D001374:Azacitidine / Q000494:pharmacology; D002294:Carcinoma, Squamous Cell / Q000235:genetics* / Q000401:mortality / Q000473:pathology; D019175:DNA Methylation / Q000187:drug effects; D004938:Esophageal Neoplasms / Q000235:genetics* / Q000401:mortality / Q000473:pathology; D000077277:Esophageal Squamous Cell Carcinoma; D005260:Female; D015972:Gene Expression Regulation, Neoplastic / Q000187:drug effects; D020868:Gene Silencing / Q000187:drug effects; D016147:Genes, Tumor Suppressor / Q000502:physiology*; D006801:Humans; D008297:Male; D008562:Membrane Glycoproteins / Q000502:physiology*; D008875:Middle Aged; D011401:Promoter Regions, Genetic / Q000187:drug effects; D014407:Tumor Cells, Cultured
chemical_list: D008562:Membrane Glycoproteins; C109148:SPINT2 protein, human; D001374:Azacitidine
keywords: Esophageal squamous cell carcinoma (ESCC); Methylation; SPINT2