Combination Analysis of Hypermethylated Wnt-Antagonist Family Genes as a Novel Epigenetic Biomarker Panel for Bladder Cancer Detection

Urakami, Shinji; Shiina, Hìroaki; Enokida, Hideki; Kawakami, Toshifumi; Kawamoto, Ken; Hirata, Hiroshi; Tanaka, Yuichiro; Kikuno, Nobuyuki; Nakagawa, Masayuki; Igawa, Mikio; Dahiya, Rajvir

doi:10.1158/1078-0432.ccr-05-2468

Cited by 160 publications

(121 citation statements)

References 26 publications

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“…We propose that the resultant downregulation of the Dkk3 gene is responsible for the activation of the Wnt/ b-catenin signaling pathway that contributes to the tumorigenesis of cervical cancer. Transcriptional inactivation of Dkk3 associated with promoter hypermethylation has been observed in cancer tissues, including acute lymphoblastic leukemia, nonsmall-cell lung cancer, prostate cancer and bladder cancer, 14,[30][31][32] indicating that the Dkk3 gene may be a frequent target for methylation and silencing in cancer.…”

Section: Discussionmentioning

confidence: 99%

Dkk3, downregulated in cervical cancer, functions as a negative regulator of β‐catenin

Lee

Rho

et al. 2008

Intl Journal of Cancer

133

136

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The Wnt/b-catenin signaling pathway is activated during the malignant transformation of keratinocytes that originate from the human uterine cervix. Dkk1, 2 and 4 have been shown to modulate the Wnt-induced stabilization of the b-catenin signaling pathway. However, the function of Dkk3 in this pathway is unknown. Comparison of the Dkk3 gene expression profiles in cervical cancer and normal cervical tissue by cDNA microarray and subsequent real-time PCR revealed that the Dkk3 gene is frequently downregulated in the cancer. Methylation studies showed that the promoter of Dkk3 was methylated in cervical cancer cell lines and 22 (31.4%) of 70 cervical cancer tissue specimens. This promoter methylation was associated with reduced expression of Dkk3 mRNA in the paired normal and tumor tissue samples. Further, the reintroduction of Dkk3 into HeLa cervical cancer cells resulted in reduced colony formation and retarded cell growth. The forced expression of Dkk3 markedly attenuated b-catenin-responsive luciferase activity in a dose-dependent manner and decreased the b-catenin levels. By utilizing a yeast two-hybrid screen, bTrCP, a negative regulator of b-catenin was identified as a novel Dkk3-interacting partner. Coexpression with bTrCP synergistically enhanced the inhibitory function of Dkk3 on b-catenin. The stable expression of Dkk3 blocks the nuclear translocation of b-catenin, resulting in downregulation of its downstream targets (VEGF and cylcin D), whereas knockdown of Dkk3 abrogates this blocking. We conclude from our finding that Dkk3 is a negative regulator of b-catenin and its downregulation contribute to an activation of the b-catenin signaling pathway.

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Section: Discussionmentioning

confidence: 99%

Dkk3, downregulated in cervical cancer, functions as a negative regulator of β‐catenin

Lee

Rho

et al. 2008

Intl Journal of Cancer

133

136

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“…34 SFRP genes are inactivated by promoter methylation in different human cancers and serve as epigenetic tumor biomarkers. [35][36][37][38] We have demonstrated that SFRP1 is a candidate TSG that is silenced in hepatocarcinogenesis through promoter hypermethylation. 39 To confirm and extend these findings with respect to SFRPs, the methylation status of other SFRP members, including SFRP2, SFRP4 and SFRP5, was also examined by MS-PCR in our study.…”

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confidence: 99%

SFRP1 suppressed hepatoma cells growth through Wnt canonical signaling pathway

Shih

Hsieh

Lai

et al. 2007

Intl Journal of Cancer

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Oncogenic activation of the Wnt/b-catenin signaling pathway is common in hepatocellular carcinoma (HCC). The secreted frizzled-related proteins (SFRPs) function as negative regulators of Wnt signaling and have important implications for carcinogenesis. Promoter hypermethylation of SFRP genes is common in human cancers. However, the role of SFRPs in HCC is not clear. Recently, we have shown that SFRP1 is frequently downregulated through promoter hypermethylation. To confirm and extend these findings, the methylation status of the other SFRP members, including SFRP2, SFRP4 and SFRP5, was examined by methylation-specific polymerase chain reaction (MS-PCR). Hypermethylation of SFRP genes, except for SFRP4, is frequent in HCCs and the levels found here were significantly higher than those seen in cirrhotic livers, chronic hepatitis livers and normal controls (p < 0.0001 for SFRP1 and SFRP2, p < 0.05 for SFRP5). To investigate the role of SFRP1 in HCCs, we used re-expression of SFRP1 in bcatenin-dependent HCC cell lines: Huh6 and HepG2. Restoration of SFRP1 attenuated Wnt signaling in those Huh6 hepatoma cells with a b-catenin gene point mutation, decreased abnormal accumulation of b-catenin in the nucleus and suppressed cell growth. Conversely, restoration of SFRP1 in HepG2 hepatoma cells with truncated bcatenin could not block the Wnt signaling pathway. Furthermore, knocking down SFRP1 by RNA interference in b-catenin-deficient cell lines (SK-Hep1) stimulated Wnt signaling and promoted cell growth. Our data suggested that SFRP1 suppressed liver cancer cells growth through Wnt canonical signaling. Moreover, b-cateninindependent noncanonical pathway might be involved in Wnt signaling activation through unknown molecules in HCC. ' 2007 Wiley-Liss, Inc.Key words: epigenetic inactivation; hepatocellular carcinoma; SFRP1; Wnt/b-catenin Hepatocellular carcinoma (HCC) is one of the most common malignant human tumors worldwide. 1-4 The major factors associated with HCC include chronic hepatitis B and C viral infections, chronic alcohol consumption, aflatoxin-B1-contaminated food and virtually all cirrhosis-inducing conditions. 4 Other etiological factors have also been proposed to lead to HCC. In addition, gender can also influence the risk and behavior of HCC, with men accounting for more cases. 5 The molecular mechanisms of hepatocarcinogenesis and the complex interactions of genetic factors remain to be elucidated.Wnt glycoproteins comprise a family of extracellular signaling ligands that play essential roles in proliferation, patterning and fate determination during normal developmental processes. [6][7][8][9][10][11][12] Although this signaling is critical for normal embryonic development, the aberrant of activation of the Wnt signal transduction pathway has been closely linked to tumorigenesis in adults. [13][14][15] b-Catenin is a crucial downstream component of the Wnt signaling pathway. When Wnt signaling is engaged, the adenomatosis polyposis coli (APC) and Axin proteins no longer bind b-catenin, with consequent b...

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“…In recent investigations, several antagonists of Wnt pathway have been identified and can be divided into two functional classes, the secreted frizzled-related protein (sFRP) class and the Dickkopf (Dkk) class [11]. As Wnt-antagonist genes, sFRP-1, sFRP-2, sFRP-4, sFRP-5, Wif-1 and Dkk-3 inhibited Wnt signaling by directly binding to Wnt molecules or by binding to the LRP5/LRP6 component of the Wnt receptor complex.…”

mentioning

confidence: 99%

“…Thus, the functional loss of Wnt antagonists can contribute to activation of the Wnt pathway and induced ectopic expression of ß-catenin. Up to now, downregulation of Wnt-antagonist genes have been identified in a variety of malignancies, including bladder [8,11,12], lung [13,14], breast [15], and chronic lymphocytic leukemia [16], and even the gastric carcinoma [17,18]and esophageal carcinoma [19]. Inactivation caused by promoter hypermethylation seems to be one of the mechanisms underlying down-regulation of the Wnt antagonists, which strongly suggests Wnt-antagonist genes function as tumor suppressor genes may play important role in tumorigenesis.…”

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confidence: 99%

Hypermethylation and aberrant expression of Wnt-antagonist family genes in gastric cardia adenocarcinoma

Guo

Guo²,

Chen³

et al. 2011

neo

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The canonical Wnt signalling pathway plays a key role during embryogenesis and pathogenesis of various types of tumors. Recently, several studies have shown that the promoter hypermethylation of Wnt-antagonist genes, including sFRP-1, , have been certified to contribute to the tumorigenesis of several cancers.The aim of this study was to investigate the promoter methylation of Wnt-antagonist genes in gastric cardia adenocarcinoma (GCA) and corresponding adjacent non-cancerous tissues, and to establish the possible relationship between DNA methylation status and the pathogenesis of GCA.MSP, RT-PCR methods were applied respectively to examine the CpG methylation of the Wnt-antagonist genes and its mRNA expression in tumors and corresponding non-cancerous tissues, and immunohistochemistry method was used to determine protein expression of β-catenin(the key factor of the Wnt signalling pathway) and cyclin D1(the target gene of this pathway).The frequency of promoter methylation of sFRP-1, sFRP-2, sFRP-4, sFRP-5, Wif-1 and Dkk-3 genes in GCA tumor tissues were 78.7%(74/94), 76.6%(72/94), 70.2%(66/94), 77.1%(73/94), 61.7%(58/94) and 21.3%(20/94), respectively, which were significantly higher than those in adjacent non-cancerous tissues. Furthermore, the frequencies of silenced mRNA expression of these six genes in GCA tumor tissues were significantly higher than those in adjacent non-cancerous tissues. Methylation levels of these six genes were all correlated with loss of mRNA expression. The ectopic expression of β-catenin and cyclin D1 was significantly more frequent in GCA tumor tissues than that in adjacent non-cancerous tissues and correlated with each Wnt-antagonist genes hypermethylation status.Epigenetic silencing of Wnt-antagonist genes expression by promoter hypermethylation may play an important role in gastric cardia adenocarcinoma.

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Combination Analysis of Hypermethylated Wnt-Antagonist Family Genes as a Novel Epigenetic Biomarker Panel for Bladder Cancer Detection

Cited by 160 publications

References 26 publications

Dkk3, downregulated in cervical cancer, functions as a negative regulator of β‐catenin

Dkk3, downregulated in cervical cancer, functions as a negative regulator of β‐catenin

SFRP1 suppressed hepatoma cells growth through Wnt canonical signaling pathway

Hypermethylation and aberrant expression of Wnt-antagonist family genes in gastric cardia adenocarcinoma

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