Eukaryotic cells maintain an immense amount of genetic information by tightly wrapping their DNA around positively charged histones. While this strategy allows human cells to maintain more than 25,000 genes, histone binding can also block gene expression.
Consequently, cells express histone acetyl transferases (HATs) to acetylate histone lysines and release DNA for transcription. Conversely, histone deacetylases (HDACs) are employed for restoring the positive charge on the histones, thereby silencing gene expression by increasing histone-DNA binding. It has previously been shown that histones bind and silence viral DNA, while hyperacetylation of histones via HDAC inhibition restores viral gene expression. In this study, we demonstrate that treatment with Entinostat, an HDAC inhibitor, enhances transgene (luciferase) expression by up to 25-fold in human prostate and murine bladder cancer cell lines when used with cationic polymers for plasmid DNA delivery. Entinostat treatment altered cell cycle progression, resulting in significant increase in the fraction of cells present in the G0/G1 phase at low micromolar concentrations. While this moderate G0/G1 arrest disappeared at higher concentrations, a modest increase in the fraction of apoptotic cells and a decrease in cell proliferation were observed, consistent with the known anticancer effects of the drug. DNase accessibility studies revealed no significant change in plasmid transcriptional availability with Entinostat treatment. However, quantitative PCR studies indicated that Entinostat treatment, at the optimal dose for enhancing transgene expression, led to an increase in the amount of plasmid present in the nucleus in two cancer cell lines. Taken together, our results show that Entinostat enhances polymer-mediated transgene expression and can be useful in applications related to transient protein expression in mammalian cells. This article is protected by copyright. All rights reserved.
Biotechnology and bioengineering. 2015 Nov 28 [Epub ahead of print]
Jacob J Elmer, Matthew D Christensen, Sutapa Barua, Jennifer Lehrman, Karmella A Haynes, Kaushal Rege
Chemical Engineering, Arizona State University, Tempe, AZ, 85287. , Chemical Engineering, Arizona State University, Tempe, AZ, 85287. , Chemical Engineering, Arizona State University, Tempe, AZ, 85287. , Harrington Biomedical Engineering, Arizona State University, Tempe, AZ, 85287. , Harrington Biomedical Engineering, Arizona State University, Tempe, AZ, 85287. , Chemical Engineering, Arizona State University, Tempe, AZ, 85287.