Histone acetyltransferase promotes fluoride toxicity in LS8 cells.
Histone acetyltransferase promotes fluoride toxicity in LS8 cells.
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组蛋白乙酰转移酶促进LS8细胞中的氟化物毒性。
DOI：
10.1016/j.chemosphere.2020.125825
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发表时间：
2020-05
期刊：
影响因子：
8.8
通讯作者：
Suzuki M
中科院分区：
文献类型：
--
作者：
Deng H;Fujiwara N;Cui H;Whitford GM;Bartlett JD;Suzuki M
先前我们证明了氟增加了来自小鼠牙釉质器官上皮和啮齿动物成釉细胞的LS8细胞中的乙酰化p53 （Ac-p53）。然而，p53如何被氟化物乙酰化以及p53上游分子途径对氟化物的反应尚不清楚。本研究表明，氟可以激活组蛋白乙酰转移酶（HATs），包括CBP、p300、PCAF和Tip60，从而使p53乙酰化。HAT活性受翻译后修饰如乙酰化和磷酸化调节。Western blots分析HAT蛋白及其翻译后修饰（p300、Acetyl-p300、CBP、Acetyl-CBP、Tip60和phospho-Tip60）。用共免疫沉淀法（co-IP）检测p53-HAT的结合。MTT法检测细胞生长抑制情况。用NaF处理LS8细胞，NaF加/不加HAT抑制剂MG149 （Tip60抑制剂）和无心酸（AA，抑制p300/CBP和PCAF）。在NaF处理前1小时加入MG149或AA。Co-IP结果显示NaF增加了p53-CBP结合和p53-PCAF结合。NaF增加活性乙酰-p300、乙酰- cbp和磷- tip60水平，表明氟激活了这些HATs。MG149可降低氟化诱导的磷酸化tip60。MG149或AA处理在24 h后逆转了氟化物诱导的细胞生长抑制。MG149或AA处理降低了氟化物诱导的p53乙酰化，抑制了caspase-3的切割、DNA损伤标记γH2AX的表达和细胞色素c释放到细胞质中。这些结果表明，通过抑制细胞生长、凋亡、DNA损伤和线粒体损伤，HATs使p53乙酰化至少在一定程度上有助于氟诱导的LS8细胞毒性。因此，HAT活性的调节可能是减轻氟对成釉细胞毒性的潜在治疗靶点。
Previously we demonstrated that fluoride increased acetylated-p53 (Ac-p53) in LS8 cells that are derived from mouse enamel organ epithelia and in rodent ameloblasts. However, how p53 is acetylated by fluoride and how the p53 upstream molecular pathway responds to fluoride is not well characterized. Here we demonstrate that fluoride activates histone acetyltransferases (HATs) including CBP, p300, PCAF and Tip60 to acetylate p53. HAT activity is regulated by post-translational modifications such as acetylation and phosphorylation. HAT proteins and their post-translational modifications (p300, Acetyl-p300, CBP, Acetyl-CBP, Tip60 and phospho-Tip60) were analyzed by Western blots. p53-HAT binding was detected by co-immunoprecipitation (co-IP). Cell growth inhibition was analyzed by MTT assays. LS8 cells were treated with NaF with/without HAT inhibitors MG149 (Tip60 inhibitor) and Anacardic Acid (AA; inhibits p300/CBP and PCAF). MG149 or AA was added 1 h prior to NaF treatment. Co-IP results showed that NaF increased p53-CBP binding and p53-PCAF binding. NaF increased active Acetyl-p300, Acetyl-CBP and phosphor-Tip60 levels, suggesting that fluoride activates these HATs. Fluoride-induced phospho-Tip60 was decreased by MG149. MG149 or AA treatment reversed fluoride-induced cell growth inhibition at 24 h. MG149 or AA treatment decreased fluoride-induced p53 acetylation to inhibit caspase-3 cleavage, DNA damage marker γH2AX expression and cytochrome-c release into the cytosol. These results suggest that acetylation of p53 by HATs contributes, at least in part, to fluoride-induced toxicity in LS8 cells via cell growth inhibition, apoptosis, DNA damage and mitochondrial damage.Modulation of HAT activity may, therefore, be a potential therapeutic target to mitigate fluoride toxicity in ameloblasts.
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影响因子：
16
作者：
Charvet C;Wissler M;Brauns-Schubert P;Wang SJ;Tang Y;Sigloch FC;Mellert H;Brandenburg M;Lindner SE;Breit B;Green DR;McMahon SB;Borner C;Gu W;Maurer U
通讯作者：
Maurer U
影响因子：
3
作者：
CHEN, LS;COUWENHOVEN, RI;SNEAD, ML
通讯作者：
SNEAD, ML
影响因子：
26.1
作者：
Green, Rivka;Lanphear, Bruce;Till, Christine
通讯作者：
Till, Christine
影响因子：
3.8
作者：
Legartova, Sona;Stixova, Lenka;Bartova, Eva
通讯作者：
Bartova, Eva
影响因子：
5.2
作者：
Reed SM;Quelle DE
通讯作者：
Quelle DE