DNA methyltransferase inhibitor-mediated apoptosis in the Wnt/beta-catenin signal pathway in a renal cell carcinoma cell line


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Konaç E., Varol N., Yilmaz A., Menevse S., Sozen S.

EXPERIMENTAL BIOLOGY AND MEDICINE, vol.238, no.9, pp.1009-1016, 2013 (SCI-Expanded) identifier identifier identifier

  • Publication Type: Article / Article
  • Volume: 238 Issue: 9
  • Publication Date: 2013
  • Doi Number: 10.1177/1535370213498984
  • Journal Name: EXPERIMENTAL BIOLOGY AND MEDICINE
  • Journal Indexes: Science Citation Index Expanded (SCI-EXPANDED), Scopus
  • Page Numbers: pp.1009-1016
  • Keywords: Wnt antagonist, beta-catenin, DNA methylation, RCC, sFRP2, GSK3 beta, apoptosis, WNT ANTAGONIST GENE, TUMOR-SUPPRESSOR, CANCER, METHYLATION
  • Gazi University Affiliated: Yes

Abstract

The Wnt signaling pathway is activated in most cancer types when Wnt antagonist genes are inactivated. Glycogen synthase kinase 3 (GSK3 beta) is an important regulator of the Wnt/beta-catenin signaling pathway. The mechanisms underlying GSK3 beta regulation of neoplastic transformation and tumor development are unclear. Studies have raised the possibility that the Wnt signaling pathway may be implicated in renal cell carcinoma (RCC). Therefore, in the present study, we hypothesize that the expression and methylation status of the secreted frizzled-related protein 2 (sFRP2) gene, one of the secreted antagonists that bind Wnt protein, and re-expression of this gene with the demethylation agent (5-aza-2'-deoxycytidine; DAC) may induce apoptosis in RCC cells. To test this hypothesis, we investigated the relationship among epigenetic inactivation of sFRP2 and p-GSK3 beta (Ser9) and other Wnt antagonists (sFRP1, DKK3, WIF-1) and apoptotic factors (Bax and Caspase3) as well as the anti-apoptotic factor BCL2. Our results indicate that DAC-mediated inhibition of DNA methylation led to a re-activation of sFRP2 expression and increased expression levels of the Wnt antagonists and apoptotic factors. In contrast, the level of beta-catenin (CTNNB1) expression decreased. The p-GSK3 beta (Ser9) protein level in Caki-2 cells was significantly down-regulated, while the DNA fragmentation rate increased after treatment with 5 mu M DAC at 96 h. Our data show that sFRP2 functions as a tumor suppressor gene in RCC and that its restoration may offer a new therapeutic approach for the treatment of RCC. Moreover, our study draws attention to the regulatory features of epigenetic molecules and analyses their underlying molecular mechanisms of action and their potential use in clinical practice.