Triagem e caracterização citotóxica e molecular de novos modelos de hidroxamatos sintéticos

Abstract

Commonly, alterations in the epigenome are observed in several tumoral lines, being able to be responsible for the development and progression of some types of cancer. This work aims to evaluate the sensitivity of in vitro cell lines to different synthetic hydroxamates, possible histone deacetylase inhibitors (HDAC). The method for evaluating cytotoxicity and selectivity was MTT. Three molecules were evaluated. In the molecular character, the in silico gene expression profiles of HDACs 1, 2, 3 and 6 were observed in two strains through the ProteinAtlasHuman platform. Additionally, the in silico primers design and in vitro validation were used to evaluate the expression of the HDAC1 and HDAC3 genes in strains treated with the chosen compound. The evaluation of the gene expression was performed using quantitative real-time PCR (qPCR) technique. The compounds tested showed a percentage of inhibition greater than 75% in the HCT116, PC3 and SF295 lines. In addition, the molecules showed lower IC 50 and selective profiles in the HL60 line, and the HDX566 compound, which presented IC 50 <4, 5 μM and selectivity index >2, in the aforementioned lineage. The HDAC1, HDAC2, HDAC3 and HDAC6 genes were analyzed in the in silico prediction model, and the HDAC1, HDAC3 and HDAC6 genes were the most expressed gene among PC3 and HL60. A parameter for the observation of the modulatory potential of these transcripts when treated with the compounds in relation to the evaluated target genes. Among the designed primers, those with the best in vitro results, which included the HDAC1 and HDAC3 genes, were chosen. Treatment with compound HDX566 in HL60 line for 24 h seems to have reduced HDAC1 gene expression. The HDAC3 gene was not expressed in any of the conditions evaluated, nor in the negative control without treatment. The present study showed three molecules with cytotoxic and selective potential. HDX566 altered the pattern of HDAC1 transcripts in the leukemic lineage evaluated.