| Chapter title |
Assessment of HDACi-Induced Cytotoxicity
|
|---|---|
| Chapter number | 3 |
| Book title |
HDAC/HAT Function Assessment and Inhibitor Development
|
| Published in |
Methods in molecular biology, January 2017
|
| DOI | 10.1007/978-1-4939-6527-4_3 |
| Pubmed ID | |
| Book ISBNs |
978-1-4939-6525-0, 978-1-4939-6527-4
|
| Authors |
Lisa Marx-Blümel, Christian Marx, Marie Kühne, Jürgen Sonnemann |
| Abstract |
The chromatin contains the genetic and the epigenetic information of a eukaryotic organism. Posttranslational modifications of histones, such as acetylation and methylation, regulate their structure and control gene expression. Histone acetyltransferases (HATs) acetylate lysine residues in histones while histone deacetylases (HDACs) remove this modification. HDAC inhibitors (HDACi) can alter gene expression patterns and induce cytotoxicity in cancer cells. Here we provide an overview of methods to determine the cytotoxic effects of HDACi treatment. Our chapter describes colorimetric methods, like trypan blue exclusion test, crystal violet staining, lactate dehydrogenase assay, MTT and Alamar Blue assays, as well as fluorogenic methods like TUNEL staining and the caspase-3/7 activity assay. Moreover, we summarize flow cytometric analysis of propidium iodide uptake, annexin V staining, cell cycle status, ROS levels, and mitochondrial membrane potential as well as detection of apoptosis by Western blot. |
Mendeley readers
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|---|---|---|
| Unknown | 4 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Student > Ph. D. Student | 1 | 25% |
| Student > Bachelor | 1 | 25% |
| Student > Postgraduate | 1 | 25% |
| Unknown | 1 | 25% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 2 | 50% |
| Biochemistry, Genetics and Molecular Biology | 1 | 25% |
| Unknown | 1 | 25% |