Reactivating aberrantly hypermethylated p15 gene in leukemic T cells by a phenylhexyl isothiocyanate mediated inter-active mechanism on DNA and chromatin.
Reactivating aberrantly hypermethylated p15 gene in leukemic T cells by a phenylhexyl isothiocyanate mediated inter-active mechanism on DNA and chromatin.
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DOI：
10.1186/1756-8722-3-48
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发表时间：
2010-11-29
影响因子：
28.5
通讯作者：
Chiao JW
中科院分区：
文献类型：
--
作者：
Jiang S;Ma X;Huang Y;Xu Y;Zheng R;Chiao JW
我们之前已经证明，苯基己基异硫氰酸酯（PHI），一种合成的异硫氰酸酯，以浓度依赖的方式抑制组蛋白去乙酰化酶和重塑染色质，诱导HL-60髓系白血病细胞的生长停滞。为了研究新型组蛋白去乙酰化酶抑制剂（HDACi） PHI对急性淋巴细胞白血病Molt-4细胞p15去甲基化和转录激活的影响，并进一步揭示其去甲基化的潜在机制，采用DNA测序和修饰甲基化特异性PCR （MSP）技术筛选了经PHI、5-Aza和TSA处理后的Molt-4细胞p15- m和p15- u mRNA。RT-PCR检测DNA甲基转移酶1 （DNMT1）、3A （DNMT3A）、3B （DNMT3B）和p15 mRNA水平。Western Blot检测P15蛋白、乙酰化组蛋白H3和组蛋白H4。Molt-4细胞中p15基因高甲基化且失活。暴露于PHI 5天后，p15基因的高甲基化减弱，p15基因以浓度依赖的方式重新激活。DNMT1和DNMT3B均受PHI抑制（P < 0.05）。在这些浓度下，DNMT3A的变化不显著。暴露于PHI后，乙酰化组蛋白H3和组蛋白H4显著积累。PHI可以诱导Molt-4细胞DNA去甲基化和乙酰化H3和H4积累。p15基因的高甲基化被逆转，p15的转录可以被PHI重新激活。
We have previously demonstrated that phenylhexyl isothiocyanate (PHI), a synthetic isothiocyanate, inhibits histone deacetylases and remodels chromatins to induce growth arrest in HL-60 myeloid leukemia cells in a concentration-dependent manner. To investigate the effect of PHI, a novel histone deacetylases inhibitor (HDACi), on demethylation and activation of transcription of p15 in acute lymphoid leukemia cell line Molt-4, and to further decipher the potential mechanism of demethylation, DNA sequencing and modified methylation specific PCR (MSP) were used to screen p15-M and p15-U mRNA after Molt-4 cells were treated with PHI, 5-Aza and TSA. DNA methyltransferase 1 (DNMT1), 3A (DNMT3A), 3B (DNMT3B) and p15 mRNA were measured by RT-PCR. P15 protein, acetylated histone H3 and histone H4 were detected by Western Blot. The gene p15 in Molt-4 cells was hypermethylated and inactive. Hypermethylation of gene p15 was attenuated and p15 gene was activated de novo after 5 days exposure to PHI in a concentration-dependent manner. DNMT1 and DNMT3B were inhibited by PHI (P < 0.05). Alteration of DNMT3A was not significant at those concentrations. Acetylated histone H3 and histone H4 were accumulated markedly after exposure to PHI. PHI could induce both DNA demethylation and acetylated H3 and H4 accumulation in Molt-4 cells. Hypermethylation of gene p15 was reversed and p15 transcription could be reactivated de novo by PHI.
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DOI：
10.1073/pnas.93.18.9821
发表时间：
1996-09-03
影响因子：
11.1
作者：
Herman, JG;Graff, JR;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
28.5
作者：
Siegel D;Hussein M;Belani C;Robert F;Galanis E;Richon VM;Garcia-Vargas J;Sanz-Rodriguez C;Rizvi S
通讯作者：
Rizvi S
影响因子：
30.8
作者：
Rountree, MR;Bachman, KE;Baylin, SB
通讯作者：
Baylin, SB
影响因子：
11.4
作者：
Melki, JR;Warnecke, P;Clark, SJ
通讯作者：
Clark, SJ
影响因子：
6.5
作者：
Tien, HF;Tang, JL;Shen, MC
通讯作者：
Shen, MC