表没食子儿茶素没食子酸酯对肺癌细胞中WIF-1启动子的去甲基化作用。
Promoter demethylation of WIF-1 by epigallocatechin-3-gallate in lung cancer cells.
作者信息
Gao Zhi, Xu Zhidong, Hung Ming-Szu, Lin Yu-Ching, Wang Tianyou, Gong Min, Zhi Xiuyi, Jablon David M, You Liang
机构信息
Department of Cardiothoracic Surgery, Beijing Friendship Hospital, Beijing, P.R.C.
出版信息
Anticancer Res. 2009 Jun;29(6):2025-30.
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 beta-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 beta-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.
背景
Wnt抑制因子-1（WIF-1）启动子异常甲基化是人类癌症表观遗传沉默的基本机制。表没食子儿茶素-3-没食子酸酯（EGCG）已被报道可直接重新激活多个甲基化沉默基因。此前尚未报道过WIF-1启动子去甲基化和重新激活的情况。
材料与方法
进行甲基化特异性PCR、测序分析和RT-PCR分析以评估WIF-1启动子去甲基化和WIF-1表达，进行蛋白质印迹分析和荧光素酶报告基因检测以评估胞质β-连环蛋白水平和Tcf/Lef报告基因活性。
结果
在H460和A549细胞系中证实了EGCG处理后WIF-1启动子去甲基化和WIF-1表达的恢复。EGCG还降低了胞质β-连环蛋白水平并抑制了Tcf/Lef报告基因活性。
结论
这些结果表明EGCG在逆转WIF-1启动子甲基化方面具有潜在的治疗用途。
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