In vitro cytotoxicity evaluation of HDAC inhibitor Apicidin in pancreatic carcinoma cells subsequent time and dose dependent treatment.

Abstract

Apicidin is a potent histone deacetylase inhibitor (HDACI) that selectively binds to histone deacetylases (HDACs) class I and interferes with the deacetylation process, which results in modification of acetylation level of cellular proteins. The aim of the study was to investigate the potential time and dose dependent cytotoxicity of the test compound, Apicidin, in pancreatic cancer cells Capan-1 and Panc-1 as well as estimate maximal tolerable dose (MTD) of the test agent and determine EC50 using four complementary colorimetric cytotoxicity or viability assays. The cells were treated with increasing concentrations of Apicidin (0-5000nM) for 2, 4 and 6h (short term exposure) or 24, 48 and 72h (long term exposure) before conducting cytotoxic analyses with lactate dehydrogenase assay or viability analyses with sulforhodamine B (SRB), methyl tetrazolium (MTT) and crystal violet (CV) assays. In order to investigate whether Apicidin irreversibly affects the cells already during the short term exposure, the medium containing Apicidin was removed and replaced with fresh culturing medium after 6h of treatment. The cells were then incubated for additional 24, 48 or 72h before carrying out the analysis. The results obtained from cytotoxicity and viability assays indicated, that Apicidin was well tolerated by both cell lines at concentrations below 100nM at any given time point and at all applied concentrations during the short term (6h or less) treatment. Continuous prolonged term exposures (48h or greater) of the cells to Apicidin with concentration exceeding 100nM resulted in significantly increasing cytotoxicity and sustained significant loss of cell viability. Moreover, long term exposure of pancreatic cancer cells Capan-1 and Panc-1 to Apicidin concentrations exceeding 100nM showed an initial anti-proliferative effect before cytotoxicity onset. In summary, MTD was exposure time dependent and estimated to 100nM for long term treatment and to at least 5000nM for treatment not greater than 6h. EC50 concentration of Apicidin was established after long term treatment, however with some variation when comparing the different assays and cell lines. Results from this study may encourage reinvestigating the capacity of potent HDACI Apicidin as an attractive agent for interfering with the deacetylation process catalyzed by HDACs for potential pancreatic cancer intervention.

• Authors: Monika Bauden, Helena Tassidis, Daniel Ansari
• Journal: Toxicology letters (Toxicol Lett) Vol. 236 Issue 1 Pg. 8-15 (Jul 02 2015) ISSN: 1879-3169 [Electronic] Netherlands
• PMID: 25917448 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)
• Copyright: Copyright © 2015 Elsevier Ireland Ltd. All rights reserved.

Chemical References

• Antineoplastic Agents
• DNA, Neoplasm
• Histone Deacetylase Inhibitors
• Mycotoxins
• Neoplasm Proteins
• Peptides, Cyclic
• apicidin
• Histone Deacetylases

Topics

• Antineoplastic Agents (pharmacology)
• Carcinoma (drug therapy, enzymology, metabolism)
• Cell Line, Tumor
• Cell Proliferation (drug effects)
• Cell Survival (drug effects)
• DNA, Neoplasm (metabolism)
• Histone Deacetylase Inhibitors (pharmacology)
• Histone Deacetylases (chemistry, metabolism)
• Humans
• Inhibitory Concentration 50
• Kinetics
• Mitochondria (drug effects, metabolism)
• Mycotoxins (pharmacology)
• Neoplasm Proteins (antagonists & inhibitors, metabolism)
• Pancreatic Neoplasms (drug therapy, enzymology, metabolism)
• Peptides, Cyclic (pharmacology)
• Reproducibility of Results