Inhibitors of histone deacetylases (HDACs) are a group of compounds displaying clear anticancer activity. They have been shown to inhibit proliferation, induce apoptosis and augment differentiation in a variety of tumour cells in vitro. Valproic acid (VPA), a well known antiepileptic drug, has been shown to inhibit HDACs in some transformed cell lines. Several studies confirmed the influence of VPA on proliferation, apoptosis and differentiation processes in malignant cells. Besides, its antitumor properties include the inhibition of angiogenesis and metastasis. In melanoma cells VPA induced the cell cycle arrest in G1 phase as well as apoptosis. This effect was associated with up-regulation of P16 protein – a cell cycle inhibitor. Generally, VPA is considered as a candidate drug useful both in the chemotherapy of advanced neoplasias and chemoprevention or control of residual minimal disease. Clinical trials included  phase I/II study of the therapy with VPA combined standard chemoimmunotherapy of patients suffering from advanced stage melanoma.
Butyric acid, a four-carbon fatty acid, is a well known inhibitor of histone deacetylases. It is formed in the human colon as a result of anaerobic bacterial fermentation of dietary fiber. It is believed that butyrate plays an important chemopreventive role in colorectal carcinogenesis.
The aim of our study was to compare the influence of VPA and sodium butyrate (NaB) on morphology, growth rate and apoptosis in human melanoma cell line A375. Both compounds were used at concentrations ranging from 0.1 to 10 mM. Cell proliferation was measured using sulforhodamine B, a protein-binding dye. Apoptosis was characterized morphologically by acridine orange staining of detached cells to visualize the condensed chromatin and fragmented nuclei of apoptotic cells. Moreover, caspase-3 activity was determined for reliable quantitative evaluation of apoptosis.
Inhibition of cell growth was found in cells treated with both compounds at the concentration of 1 mM. Incubation of cells with tested compounds at concentrations of 3mM and 10mM resulted in considerable changes in cell morphology. Numerous cells detached from the tissue culture dishes and floated in the medium. Examination of these cells by acridine orange staining revealed the typical morphological characteristics of apoptosis such as condensed chromatin and fragmented nuclei. The manner of action of examined compounds (VPA and NaB) was similar.

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