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.

ZNF671 DNA methylation as a molecular predictor for the early recurrence of serous ovarian cancer

“…ZNF671 contains Krüppel‐associated box (KRAB) and C 2 H 2 ‐type zinc finger domains and functions as a transcription factor, although the precise roles of this protein have not been fully elucidated. In previous reports, ZNF671 functioned as a tumor suppressor in urothelial cancer and nasopharyngeal cancer . In addition, the hypermethylation of ZNF671 was observed in several cancers, such as clear cell renal carcinomas and urothelial cancers .…”

“…In addition, the hypermethylation of ZNF671 was observed in several cancers, such as clear cell renal carcinomas and urothelial cancers . In particular, the DNA methylation of ZNF671 was reported to be a potential biomarker for the relapse of disease in urothelial cancer . In ovarian cancer, ZNF671 is 1 of 168 genes whose expressions are silenced by DNA methylation in clinical cases .…”

Is Overexpression of Ki-67 a Prognostic Biomarker of Upper Tract Urinary Carcinoma? A Retrospective Cohort Study and Meta-Analysis

“…Upper tract urinary carcinoma (UTUC) is a relatively uncommon but aggressive disease that accounts for only 5-10% of all urothelial malignancies [1,2] and has an estimated annual incidence in Western countries of approximately 2 cases per 100,000 inhabitants [3]. As previously reported, the ratio of urothelial carcinoma in the renal pelvis and ureter is reported to be 3:1 [4,5].…”

Genetic and Epigenetic Alterations in Bladder Cancer

“…Yu et al [63] reported 92% sensitivity and 87% specificity for both primary and recurrent cases by monitoring DNA methylation of 11 genes. The detection sensitivity of a 3-gene panel comprised of ZNF671, IRF8 , and SFRP1 DNA methylation was 96% and approached 100% for high-grade and recurrent patients, compared to only 58% sensitivity using DNA methylation of ZNF671 alone [64]. DNA methylation of TWIST and NID2 yielded 90% sensitivity and 96% specificity for predicting bladder cancer recurrence [65].…”