Histone Deacetylase Inhibition and Autophagy Modulation Induces a Synergistic Antiproliferative Effect and Cell Death in Cholangiocarcinoma Cells.

Cholangiocarcinoma, also known as biliary tract cancer, is an aggressive adenocarcinoma arising from epithelial cells lining the intra- and extrahepatic biliary system. The effects of autophagy modulators and histone deacetylase (HDAC) inhibitors in cholangiocarcinoma are not fully known. It is essential to understand the molecular mechanisms and the effects of HDAC inhibitors in the context of cholangiocarcinoma. The antiproliferative effect of different HDAC inhibitors and autophagy modulation was investigated by the MTT cell viability assay in TFK-1 and EGI-1 cholangiocarcinoma cell lines. Combination indexes were calculated using CompuSyn software. Consequently, apoptosis was detected by Annexin V/PI staining. The effect of the drugs on the cell cycle was measured by the propidium iodide staining. The HDAC inhibition was confirmed via acetylated histone protein levels by western blotting. HDAC inhibitors, MS-275 and romidepsin, showed a better synergistic effect with the nocodazole combination. The combination treatment exerted its growth inhibitory effect by cell cycle arrest and induction of apoptosis. The cell cycle analysis of the combination treatment showed that the S phase and G2/M phase were achieved. Moreover, the necrotic and apoptotic cell population increased after single HDAC inhibitors and combination treatment. The anti-cancer effect of HDAC inhibitors is revealed by acetylation levels of histones. While acetylation levels were increased in response to HDAC inhibitors and autophagy modulator combinations, the HDAC expression decreased. This study highlights the importance of the combination of HDAC inhibition and autophagy modulators and demonstrates a synergistic effect, which could be a promising therapy and novel treatment approach for cholangiocarcinoma.