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

Melina Mitsiogianni,Dimitrios T. Trafalis,Aglaia Pappa,Mihalis I. Panayiotidis,Sotiris Botaitis,Vasilis Zoumpourlis,H. P. Vasantha Rupasinghe,Theodora Mantso,Rodrigo Franco

doi:10.1007/s00394-019-01925-6

Abstract

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

Highlights

Melanoma is an aggressive and highly metastatic type of skin cancer with significantly increasing incidence rates over the past few years [1, 2]
It is evident that allyl isothiocyanate (AITC)-induced reduction in cell viability cannot be linked to an increased DNA methylation status as the co-treatment protocol did not reverse the cytotoxic effect of AITC in A375 cells
Our results showed that exposure to AITC (2.5–50 μM) reduced viability in human A375 and Hs 294T as well as murine B16-F10 melanoma cells in a concentration- and time-dependent manner

Summary

Introduction

Melanoma is an aggressive and highly metastatic type of skin cancer with significantly increasing incidence rates over the past few years [1, 2]. The design of new approaches in disease prevention and treatment is of great importance To this end, consumption of cruciferous vegetables has been strongly associated with reduced risk of cancer development because of their rich content in isothiocyanates (ITCs) [3]. Consumption of cruciferous vegetables has been strongly associated with reduced risk of cancer development because of their rich content in isothiocyanates (ITCs) [3] These are compounds produced through hydrolysis of their precursor molecules, glucosinolates, by an enzyme called myrosinase which is activated after plant tissue disruption [4]. Further evidence supports the involvement of ITCs in gene regulation by reversing cancer-associated epigenetic marks at both DNA and histone levels [16,17,18,19,20,21]

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