title: Epigenetic silencing of the NR4A3 tumor suppressor, by aberrant JAK/STAT signaling, predicts prognosis in gastric cancer.
abstract: While aberrant JAK/STAT signaling is crucial to the development of gastric cancer (GC), its effects on epigenetic alterations of its transcriptional targets remains unclear. In this study, by expression microarrays coupled with bioinformatic analyses, we identified a putative STAT3 target gene, NR4A3 that was downregulated in MKN28 GC daughter cells overexpressing a constitutively activated STAT3 mutant (S16), as compared to an empty vector control (C9). Bisulphite pyrosequencing and demethylation treatment showed that NR4A3 was epigenetically silenced by promoter DNA methylation in S16 and other GC cell lines including AGS cells, showing constitutive activation of STAT3. Subsequent experiments revealed that NR4A3 promoter binding by STAT3 might repress its transcription. Long-term depletion of STAT3 derepressed NR4A3 expression, by promoter demethylation, in AGS GC cells. NR4A3 re-expression in GC cell lines sensitized the cells to cisplatin, and inhibited tumor growth in vitro and in vivo, in an animal model. Clinically, GC patients with high NR4A3 methylation, or lower NR4A3 protein expression, had significantly shorter overall survival. Intriguingly, STAT3 activation significantly associated only with NR4A3 methylation in low-stage patient samples. Taken together, aberrant JAK/STAT3 signaling epigenetically silences a potential tumor suppressor, NR4A3, in gastric cancer, plausibly representing a reliable biomarker for gastric cancer prognosis.
authors.affiliation: Department of Surgical Pathology, Changhua Christian Hospital, Changhua, Taiwan.|Department of Medical Technology, Jen-Teh Junior College of Medicine, Nursing and Management, Miaoli, Taiwan.
authors.forename: Chung-Min
authors.initials: CM
authors.lastname: Yeh
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Liang-Yu
authors.initials: LY
authors.lastname: Chang
authors.affiliation: Department of Surgical Pathology, Changhua Christian Hospital, Changhua, Taiwan.|Department of Medical Technology, Jen-Teh Junior College of Medicine, Nursing and Management, Miaoli, Taiwan.|Institute of Medicine, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Shu-Hui
authors.initials: SH
authors.lastname: Lin
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Jian-Liang
authors.initials: JL
authors.lastname: Chou
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Department of Medical Research, Ditmanson Medical Foundation Chiayi Christian Hospital, Chia-Yi, Taiwan.
authors.forename: Hsiao-Yen
authors.initials: HY
authors.lastname: Hsieh
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Li-Han
authors.initials: LH
authors.lastname: Zeng
authors.affiliation: Department of Computer Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Sheng-Yu
authors.initials: SY
authors.lastname: Chuang
authors.affiliation: Department of Computer Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Hsiao-Wen
authors.initials: HW
authors.lastname: Wang
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Zentrum für Molekulare Biologie Heidelberg (ZMBH), DKFZ-ZMBH Alliance, Heidelberg, Germany.
authors.forename: Claudia
authors.initials: C
authors.lastname: Dittner
authors.affiliation: Department of Surgical Pathology, Changhua Christian Hospital, Changhua, Taiwan.
authors.forename: Cheng-Yu
authors.initials: CY
authors.lastname: Lin
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Jora M J
authors.initials: JM
authors.lastname: Lin
authors.affiliation: Department of Computer Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Yao-Ting
authors.initials: YT
authors.lastname: Huang
authors.affiliation: Department of Surgery, The Chinese University of Hong Kong, Hong Kong SAR, China.
authors.forename: Enders K W
authors.initials: EK
authors.lastname: Ng
authors.affiliation: School of Biomedical Sciences, The Chinese University of Hong Kong, Hong Kong SAR, China.
authors.forename: Alfred S L
authors.initials: AS
authors.lastname: Cheng
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Shu-Fen
authors.initials: SF
authors.lastname: Wu
authors.affiliation: Department of Pediatrics, The Ohio State University, Columbus, OH, USA.
authors.forename: Jiayuh
authors.initials: J
authors.lastname: Lin
authors.affiliation: Department of Surgical Pathology, Changhua Christian Hospital, Changhua, Taiwan.|School of Medicine, Chung Shan Medical University, Taichung, Taiwan.
authors.forename: Kun-Tu
authors.initials: KT
authors.lastname: Yeh
authors.affiliation: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors.forename: Michael W Y
authors.initials: MW
authors.lastname: Chan
mesh_terms: D045744:Cell Line, Tumor; D019175:DNA Methylation; D004268:DNA-Binding Proteins / Q000235:genetics*; D044127:Epigenesis, Genetic; D020868:Gene Silencing; D006801:Humans; D053612:Janus Kinases / Q000378:metabolism*; D011379:Prognosis; D011401:Promoter Regions, Genetic; D011987:Receptors, Steroid / Q000235:genetics*; D011988:Receptors, Thyroid Hormone / Q000235:genetics*; D050796:STAT3 Transcription Factor / Q000378:metabolism*; D015398:Signal Transduction; D013274:Stomach Neoplasms / Q000235:genetics / Q000473:pathology*
chemical_list: D004268:DNA-Binding Proteins; C490412:NR4A3 protein, human; D011987:Receptors, Steroid; D011988:Receptors, Thyroid Hormone; D050796:STAT3 Transcription Factor; D053612:Janus Kinases