Abstract
Background: Aberrant promoter methylation of Wnt inhibitory factor-1 (WIF-1) is a fundamental mechanism of epigenetic silencing in human cancers. Epigallocatechin-3-gallate (EGCG) has been reported to directly reactivate several methylation-silenced genes. The promoter demethylation and reactivation of WIF-1 has not previously been reported. Materials and Methods: Methylation-specific PCR, sequencing analysis and RT-PCR analysis were performed to evaluate promoter demethylation of WIF-1 and WIF-1 expression, Western blot analysis and luciferase reporter assay were performed to evaluate expression of cytosolic β-catenin protein and Tcf/Lef reporter activity. Results: Promoter demethylation of WIF-1 and restoration of WIF-1 expression after EGCG treatment are demonstrated in H460 and A549 cell lines. EGCG also decreased cytosolic β-catenin protein level and inhibited Tcf/Lef reporter activity. Conclusion: These results suggest the potential therapeutic use of EGCG for the reversal of WIF-1 promoter methylation.
- Wnt pathway
- Wnt inhibitory factor-1
- promoter hypermethylation
- demethylation
- epigallocatechin-3-gallate
- EGCG
Footnotes
- Received January 29, 2009.
- Revision received March 27, 2009.
- Accepted April 15, 2009.
- Copyright© 2009 International Institute of Anticancer Research (Dr. John G. Delinassios), All rights reserved