Melina Mitsiogianni1, Dimitrios T Trafalis2, Rodrigo Franco3,4, Vasilis Zoumpourlis5, Aglaia Pappa6, Mihalis I Panayiotidis7,8. 1. Faculty of Health and Life Sciences, Department of Applied Sciences, Group of Translational Biosciences, Northumbria University, Newcastle Upon Tyne, NE1 8ST, UK. 2. Laboratory of Pharmacology, Clinical Pharmacology Unit, Medical School, National and Kapodistrian University of Athens, 11527, Athens, Greece. 3. Redox Biology Centre, University of Nebraska-Lincoln, Lincoln, NE, 68588, USA. 4. School of Veterinary Medicine & Biomedical Sciences, University of Nebraska-Lincoln, Lincoln, NE, 68583, USA. 5. Institute of Biology, Medicinal Chemistry and Biotechnology, National Hellenic Research Foundation, 11635, Athens, Greece. 6. Department of Molecular Biology and Genetics, Democritus University of Thrace, 68100, Alexandroupolis, Greece. 7. Faculty of Health and Life Sciences, Department of Applied Sciences, Group of Translational Biosciences, Northumbria University, Newcastle Upon Tyne, NE1 8ST, UK. mihalisp@cing.ac.cy. 8. Department of Electron Microscopy and Molecular Pathology, The Cyprus Institute of Neurology and Genetics, 2371, Nicosia, Cyprus. mihalisp@cing.ac.cy.
Abstract
OBJECTIVE(S): Growing evidence supports that isothiocyanates exert a wide range of bioactivities amongst of which is their capacity to interact with the epigenetic machinery in various cancers including melanoma. Our aim was to characterise the effect of sulforaphane and iberin on histone acetylation and methylation as a potential anti-melanoma strategy. METHODS: We have utilised an in vitro model of malignant melanoma [consisting of human (A375, Hs294T, VMM1) and murine (B16F-10) melanoma cell lines as well as a non-melanoma (A431) and a non-tumorigenic immortalised keratinocyte (HaCaT) cell line] exposed to sulforaphane or iberin. Cell viability was evaluated by the Alamar blue assay whilst total histone deacetylases and acetyltransferases activities were determined by the Epigenase HDAC Activity/Inhibition and EpiQuik HAT Activity/Inhibition assay kits, respectively. The expression levels of specific histone deacetylases and acetyltransferases together with those of lysine acetylation and methylation marks were obtained by western immunoblotting. RESULTS: Overall, both sulforaphane and iberin were able to (1) reduce cell viability, (2) decrease total histone deacetylase activity and (3) modulate the expression levels of various histone deacetylases as well as acetyl and methyl transferases thus modulating the acetylation and methylation status of specific lysine residues on histones 3 and 4 in malignant melanoma cells. CONCLUSIONS: Our findings highlight novel insights as to how sulforaphane and iberin differentially regulate the epigenetic response in ways compatible with their anticancer action in malignant melanoma.
OBJECTIVE(S): Growing evidence supports that isothiocyanates exert a wide range of bioactivities amongst of which is their capacity to interact with the epigenetic machinery in various cancers including melanoma. Our aim was to characterise the effect of sulforaphane and iberin on histone acetylation and methylation as a potential anti-melanoma strategy. METHODS: We have utilised an in vitro model of malignant melanoma [consisting of human (A375, Hs294T, VMM1) and murine (B16F-10) melanoma cell lines as well as a non-melanoma (A431) and a non-tumorigenic immortalised keratinocyte (HaCaT) cell line] exposed to sulforaphane or iberin. Cell viability was evaluated by the Alamar blue assay whilst total histone deacetylases and acetyltransferases activities were determined by the Epigenase HDAC Activity/Inhibition and EpiQuik HAT Activity/Inhibition assay kits, respectively. The expression levels of specific histone deacetylases and acetyltransferases together with those of lysine acetylation and methylation marks were obtained by western immunoblotting. RESULTS: Overall, both sulforaphane and iberin were able to (1) reduce cell viability, (2) decrease total histone deacetylase activity and (3) modulate the expression levels of various histone deacetylases as well as acetyl and methyl transferases thus modulating the acetylation and methylation status of specific lysine residues on histones 3 and 4 in malignant melanoma cells. CONCLUSIONS: Our findings highlight novel insights as to how sulforaphane and iberin differentially regulate the epigenetic response in ways compatible with their anticancer action in malignant melanoma.
Authors: Javier Martinez-Useros; Mario Martin-Galan; Maria Florez-Cespedes; Jesus Garcia-Foncillas Journal: Cancers (Basel) Date: 2021-06-27 Impact factor: 6.639