Effects of DNMT and HDAC Inhibitors (RG108 and Trichostatin A) on NGF-induced Neurite Outgrowth and Cellular Migration

Abstract

Neuropathological characteristics of neurodegenerative disorders are atrophy or loss of specific neurons in the specific brain areas. Small-molecule products like epigenetic drugs, which can activate silenced genes, regulate chromatin remodelling and transcription, is presently been focused as an alternative approach for eliciting neuritogenic activity. Trichostatin A (TSA) has been effectively investigated for cancer treatment but it also has been shown to possess neuritogenic potential. In this study, it was aimed to compare the neuritogenic effects of two epigenetic drugs on PC-12 Adh cell line, which are the histone deacetylase inhibitor (HDACi) trichostatin A and a novel non-covalent DNA methyltransferase inhibitor (DNMTi) RG108. Cytotoxic effects were determined by MTT assay. Cell differentiation and migration analysis were evaluated by xCELLigence Real Time Cell Analyzer Dual Plate (RTCA DP) system according to the changing in Cell Index (CI) values. Cell migration analysis was also corroborated with morphological migration analysis. Matrix metalloproteinase-2 (MMP-2) levels related with neurite outgrowth were measured by ELISA. Finally, neurite outgrowth was observed with immunofluorescence staining and determined by neurite outgrowth analysis on PC-12 Adh cell line. The RG108 was found nearly as effective as TSA on neuronal differentiation, neurite outgrowth and cellular migration in PC-12 Adh cell line in a combination with nerve growth factor. According to the results, DNA methyltransferase inhibitors also hold promise in neurodegenerative disorders.