title: Methylomics analysis identifies ZNF671 as an epigenetically repressed novel tumor suppressor and a potential non-invasive biomarker for the detection of urothelial carcinoma.
abstract: The molecular mechanism underlying the lethal phenomenon of urothelial carcinoma (UC) tumor recurrence remains unresolved. Here, by methylation microarray, we identified promoter methylation of the zinc-finger protein gene, ZNF671 in bladder UC tumor tissue samples, a finding that was independently validated by bisulphite pyrosequencing in cell lines and tissue samples. Subsequent assays including treatment with epigenetic depressive agents and in vitro methylation showed ZNF671 methylation to result in its transcriptional repression. ZNF671 re-expression in UC cell lines, via ectopic expression, inhibited tumor growth and invasion, in possible conjunction with downregulation of cancer stem cell markers (c-KIT, NANOG, OCT4). Clinically, high ZNF671 methylation in UC tumor tissues (n=96; 63 bladder, 33 upper urinary tract) associated with tumor grade and poor locoregional disease-free survival. Quantitative MSP analysis in a training (n=97) and test (n=61) sets of voided urine samples from bladder UC patients revealed a sensitivity and specificity of 42%-48% and 89%-92.8%, respectively, for UC cancer detection. Moreover, combining DNA methylation of ZNF671 and 2 other genes (IRF8 and sFRP1) further increased the sensitivity to 96.2%, suggesting a possible three-gene UC biomarker. In summary, ZNF671, an epigenetically silenced novel tumor suppressor, represents a potential predictor for UC relapse and non-invasive biomarker that could assist in UC clinical decision-making.
authors: 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: Chia-Ming
authors: CM
authors: Yeh
authors: Department of Urology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Pi-Che
authors: PC
authors: Chen
authors: Institute of Molecular Biology, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Department of Medical Research, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Hsiao-Yen
authors: HY
authors: Hsieh
authors: Department of Urology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Yeong-Chin
authors: YC
authors: Jou
authors: Department of Urology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Chang-Te
authors: CT
authors: Lin
authors: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.
authors: Ming-Hsuan
authors: MH
authors: Tsai
authors: 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: Wen-Yu
authors: WY
authors: Huang
authors: 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: Yi-Ting
authors: YT
authors: Wang
authors: Department of Life Science, National Chung Cheng University, Min-Hsiung, Chia-Yi, Taiwan.|Departments of Radiation Oncology, Buddhist Dalin Tzu Chi General Hospital, Chia Yi, Taiwan.
authors: Ru-Inn
authors: RI
authors: Lin
authors: 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: Szu-Shan
authors: SS
authors: Chen
authors: Department of Pathology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Chun-Liang
authors: CL
authors: Tung
authors: 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: Shu-Fen
authors: SF
authors: Wu
authors: 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: D Ching
authors: DC
authors: Chang
authors: Department of Urology, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Cheng-Huang
authors: CH
authors: Shen
authors: Department of Medical Research, Ditmanson Medical Foundation Chia-Yi Christian Hospital, Chia-Yi, Taiwan.
authors: Cheng-Da
authors: CD
authors: Hsu
authors: 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: Michael W Y
authors: MW
authors: Chan
mesh_terms: D000818:Animals; D014408:Biomarkers, Tumor / Q000235:genetics* / Q000652:urine; D002295:Carcinoma, Transitional Cell / Q000235:genetics* / Q000473:pathology / Q000652:urine; D045744:Cell Line, Tumor; D019175:DNA Methylation; D004273:DNA, Neoplasm / Q000737:chemistry / Q000235:genetics / Q000652:urine; D044127:Epigenesis, Genetic; D005260:Female; D015972:Gene Expression Regulation, Neoplastic; D006801:Humans; D008297:Male; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D008875:Middle Aged; D020133:Reverse Transcriptase Polymerase Chain Reaction; D017422:Sequence Analysis, DNA; D014183:Transplantation, Heterologous; D047368:Tumor Burden / Q000235:genetics; D025521:Tumor Suppressor Proteins / Q000235:genetics*; D001749:Urinary Bladder Neoplasms / Q000235:genetics* / Q000473:pathology / Q000652:urine
chemical_list: D014408:Biomarkers, Tumor; D004273:DNA, Neoplasm; D025521:Tumor Suppressor Proteins; C000609488:ZNF671 protein, human
keywords: DNA methylation; ZNF671; urine; urothelial carcinoma