丙烯基异硫氰酸酯调节恶性黑素瘤实验模型中的赖氨酸乙酰化和甲基化标记。

Allyl isothiocyanate regulates lysine acetylation and methylation marks in an experimental model of malignant melanoma.

机构信息

Department of Applied Sciences, Translational Biosciences Group, Northumbria University, Ellison Building, Newcastle Upon Tyne, NE1 8ST, UK.

Laboratory of Pharmacology, Clinical Pharmacology Unit, Medical School, National and Kapodistrian University of Athens, 11527, Athens, Greece.

出版信息

Eur J Nutr. 2020 Mar;59(2):557-569. doi: 10.1007/s00394-019-01925-6. Epub 2019 Feb 14.

OBJECTIVE(S): Isothiocyanates (ITCs) are biologically active plant secondary metabolites capable of mediating various biological effects including modulation of the epigenome. Our aim was to characterize the effect of allyl isothiocyanate (AITC) on lysine acetylation and methylation marks as a potential epigenetic-induced anti-melanoma strategy.

METHODS

Our malignant melanoma model consisted of (1) human (A375) and murine (B16-F10) malignant melanoma as well as of human; (2) brain (VMM1) and lymph node (Hs 294T) metastatic melanoma; (3) non-melanoma epidermoid carcinoma (A431) and (4) immortalized keratinocyte (HaCaT) cells subjected to AITC. Cell viability, histone deacetylases (HDACs) and acetyltransferases (HATs) activities were evaluated by the Alamar blue, Epigenase HDAC Activity/Inhibition and EpiQuik HAT Activity/Inhibition assay kits, respectively, while their expression levels together with those of lysine acetylation and methylation marks by western immunoblotting. Finally, apoptotic gene expression was assessed by an RT-PCR-based gene expression profiling methodology.

RESULTS

AITC reduces cell viability, decreases HDACs and HATs activities and causes changes in protein expression levels of various HDACs, HATs, and histone methyl transferases (HMTs) all of which have a profound effect on specific lysine acetylation and methylation marks. Moreover, AITC regulates the expression of a number of genes participating in various apoptotic cascades thus indicating its involvement in apoptotic induction.

CONCLUSIONS

AITC exerts a potent epigenetic effect suggesting its potential involvement as a promising epigenetic-induced bioactive for the treatment of malignant melanoma.

目的（S）：异硫氰酸酯（ITCs）是生物活性的植物次生代谢物，能够介导各种生物学效应，包括表观基因组的调节。我们的目的是描述丙烯基异硫氰酸酯（AITC）对赖氨酸乙酰化和甲基化标记的影响，作为一种潜在的表观遗传诱导抗黑色素瘤策略。

方法

我们的恶性黑色素瘤模型包括（1）人（A375）和鼠（B16-F10）恶性黑色素瘤，以及人；（2）脑（VMM1）和淋巴结（Hs 294T）转移性黑色素瘤；（3）非黑色素瘤表皮癌（A431）和（4）永生化角质形成细胞（HaCaT）细胞用 AITC 处理。通过 Alamar blue、Epigenase HDAC Activity/Inhibition 和 EpiQuik HAT Activity/Inhibition 测定试剂盒分别评估细胞活力、组蛋白去乙酰化酶（HDACs）和乙酰转移酶（HATs）的活性，而通过 Western 免疫印迹法评估其表达水平以及赖氨酸乙酰化和甲基化标记的表达水平。最后，通过基于 RT-PCR 的基因表达谱分析方法评估凋亡基因的表达。