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Flow Epigenetics in Cancer

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  2. Flow Epigenetics in Cancer
  3. FACS for isolating cells from patient samples

The epigenetic regulation of gene expression through specific modifications of core histone proteins plays a major role during normal development. It is now recognized that enzymes that effect these histone modifications (mainly methylation and acetylation of specific lysine sites) are frequently deranged during cancer development, and can result in the inappropriate silencing of critical tumor suppressor genes. Consequently, there is current interest in the development of novel agents that selectively target the epigenome in order to control cancer growth or restore sensitivity to standard agents. In this paper, Watson and coworkers demonstrate the potential to measure multiple histone modification sites in single cells by flow cytometry, and show that this approach can be used to monitor the effects of novel drugs to target the epigenome and to track changes within different cell populations.

In this issue: page 78

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FACS for isolating cells from patient samples

  1. Top of page
  2. Flow Epigenetics in Cancer
  3. FACS for isolating cells from patient samples

Transcriptome analysis of patient samples has become exceedingly popular in many scientific disciplines aiming to understand disease pathogenesis or responses to therapy. Minimization of the secondary effects associated with cell isolation on gene expression is crucial to successful disease or treatment-relevant transcriptome analyses. These secondary effects include exposure to cold temperature during incubations and binding to receptor-specific antibodies. Beliakova-Bethell,Massanella, and coworkers demonstrate that fluorescence activated cell sorting (FACS) is the optimal method of isolation of leukocyte subsets from total peripheral blood mononuclear cells. FACS not only results in the best purity, ensuring uncontaminated gene expression signatures, but has minimal impact on cellular transcription compared to immunomagnetic negative and positive selection. Remarkably, the stresses associated with passing cells through the flow cytometer do not appear to result in changes in gene expression. Moreover, FACS allows isolation of multiple cell subsets from the same sample, posing an additional benefit when working with precious patient samples.

In this issue: page 94

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