Targeted TET oxidase activity through methyl-CpG-binding domain extensively suppresses cancer cell proliferation.
Targeted TET oxidase activity through methyl-CpG-binding domain extensively suppresses cancer cell proliferation.
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DOI：
10.1002/cam4.830
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发表时间：
2016-09
期刊：
影响因子：
4
通讯作者：
Fukushige, Shinichi
中科院分区：
文献类型：
--
作者：
Mizuguchi, Yasuhiko;Saiki, Yuriko;Horii, Akira;Fukushige, Shinichi
关键词：
DNA甲基转移酶（DNMT）抑制剂是用于治疗骨髓增生异常综合征的表观遗传药物。它们不仅诱导DNA去甲基化，而且具有显著的细胞抑制和细胞毒性作用;然而，由于缺乏仅诱导DNA去甲基化的方法，这些特征之间的关系尚未建立。在本文中，我们发现由甲基-CpG-结合结构域（MBD）和10 - 11易位蛋白1（TET 1-CD）的催化结构域组成的融合蛋白全面去甲基化和上调许多甲基化基因。这些上调的基因通常在转录起始位点（TSS）± 1000 bp内含有CpG岛（CGI）。有趣的是，65%被MBD-TET 1-CDwt上调5倍或更多的基因在用DNMT抑制剂5-氮杂胞苷（Aza-CR）处理后也被重新激活，这表明通过两种方法进行的基因重新激活主要具有相同的机制，即DNA去甲基化。为了研究DNA去甲基化是否影响癌细胞的生长，我们建立了四环素诱导系统，该系统可以调节前列腺癌细胞系LNCaP中MBD-TET 1-CDwt的表达。MBD-TET 1-CDwt的诱导使谷胱甘肽S-转移酶pi 1（GSTP 1）去甲基化和上调，GSTP 1是前列腺癌中的高甲基化基因之一。与甲基化基因的再活化一致，MBD-TET 1-CDwt的诱导通过G1/S停滞广泛抑制LNCaP细胞的生长。这些结果清楚地表明，通过MBD在甲基-CpG位点募集的泰特氧化酶活性通过DNA去甲基化诱导高甲基化基因的再活化，并使我们能够分析各种癌细胞中仅整体DNA去甲基化的影响。
DNA methyltransferase (DNMT) inhibitors are epigenetic drugs used to treat myelodysplastic syndrome. They not only induce DNA demethylation but also have significant cytostatic and cytotoxic effects; however, the relationships between these characteristics have not been established yet due to the lack of a method to induce only DNA demethylation. Herein, we show that a fusion protein comprised of the methyl‐CpG‐binding domain (MBD) and the catalytic domain of Ten‐eleven translocation protein 1 (TET1‐CD) globally demethylates and upregulates a number of methylated genes. These upregulated genes frequently contained CpG islands (CGIs) within ± 1000 bp of the transcription start site (TSS). Interestingly, 65% of the genes upregulated fivefold or more by MBD‐TET1‐CDwt were also reactivated after treatment with a DNMT inhibitor, 5‐azacytidine (Aza‐CR), suggesting that gene reactivation by both methods primarily shares the same mechanism, DNA demethylation. In order to examine whether DNA demethylation affects the growth of cancer cells, we have established a tetracycline inducible system that can regulate the expression of MBD‐TET1‐CDwt in a prostate cancer cell line, LNCaP. The induction of MBD‐TET1‐CDwt demethylated and upregulated glutathione S‐transferase pi 1 (GSTP1), one of the hypermethylated genes in prostate cancer. In accordance with the reactivation of methylated genes, induction of MBD‐TET1‐CDwt extensively suppressed the growth of LNCaP cells through G1/S arrest. These results clearly indicate that TET oxidase activity recruited at methyl‐CpG sites through MBD induces reactivation of hypermethylated genes by DNA demethylation and allows us to analyze the effect of only global DNA demethylation in a wide variety of cancer cells.
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影响因子：
46.9
作者：
通讯作者：
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影响因子：
30.8
作者：
Rountree, MR;Bachman, KE;Baylin, SB
通讯作者：
Baylin, SB
DOI：
10.1016/j.bbrc.2009.01.010
发表时间：
2009-02-20
影响因子：
3.1
作者：
Fukushige, Shinichi;Kondo, Emiko;Horii, Akira
通讯作者：
Horii, Akira
影响因子：
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通讯作者：
Tajima S
影响因子：
4.8
作者：
Bachman, KE;Rountree, MR;Baylin, SB
通讯作者：
Baylin, SB