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New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry

The imaging analytical capabilities of laser scanning cytometer (LSC) have been used to assess morphological features considered to be typical of the senescent phenotype. The characteristic “flattening” of senescent cells was reflected by the decline in the density of staining (intensity of maximal...

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Autores principales: Zhao, Hong, Halicka, H Dorota, Traganos, Frank, Jorgensen, Ellen, Darzynkiewicz, Zbigniew
Formato: Texto
Lenguaje:English
Publicado: Wiley Subscription Services, Inc., A Wiley Company 2010
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2977923/
https://www.ncbi.nlm.nih.gov/pubmed/20939035
http://dx.doi.org/10.1002/cyto.a.20983
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author Zhao, Hong
Halicka, H Dorota
Traganos, Frank
Jorgensen, Ellen
Darzynkiewicz, Zbigniew
author_facet Zhao, Hong
Halicka, H Dorota
Traganos, Frank
Jorgensen, Ellen
Darzynkiewicz, Zbigniew
author_sort Zhao, Hong
collection PubMed
description The imaging analytical capabilities of laser scanning cytometer (LSC) have been used to assess morphological features considered to be typical of the senescent phenotype. The characteristic “flattening” of senescent cells was reflected by the decline in the density of staining (intensity of maximal pixel) of DNA-associated fluorescence [4,6-diamidino-2-phenylindole (DAPI)] paralleled by an increase in nuclear size (area). The decrease in ratio of maximal pixel to nuclear area was even more sensitive senescence biomarker than the change in maximal pixel or nuclear area, each alone. The saturation cell density at plateau phase of growth recorded by LSC was found to be dramatically decreased in cultures of senescent cells, thereby also serving as an additional marker. The induction of cyclin dependent kinase inhibitors p21(WAF1) and p27(KIP1) and γH2AX and activation of ATM markers of DNA damage response were measured in parallel with DNA/DAPI maximal pixel and nuclear area. These biomarker indices were expressed in quantitative terms by reporting them as a fraction of the respective controls. The effect of treatment of A549 and WI-38 cells with different concentrations of mitoxantrone (Mxt) and trichostatin A for various time periods was studied to assess the degree (depth) of cell senescence. Also assessed was the effect of 2-deoxy-d-glucose, the agent attenuating metabolic cell activity, on the depth of senescence induced by Mxt. A relationship between the ability of cells to synthesize RNA (incorporate 5-ethynyluridine) that leads to growth imbalance and induction of cell senescence was also studied. The data show that morphometric analysis of cellular attributes by LSC offers an attractive tool to detect cell senescence and measure its degree particularly in assessing effects of the factors that enhance or attenuate this process. This methodology is of importance in light of the evidence that cellular senescence is not only a biological process that is fundamental for organismal aging but also impedes formation of induced-pluripotent stem cells providing the barrier for neoplastic transformation and is the major mechanism of induction of reproductive cell death during treatment of solid tumors. © 2010 International Society for Advancement of Cytometry.
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spelling pubmed-29779232011-02-15 New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry Zhao, Hong Halicka, H Dorota Traganos, Frank Jorgensen, Ellen Darzynkiewicz, Zbigniew Cytometry A Rapid Publication The imaging analytical capabilities of laser scanning cytometer (LSC) have been used to assess morphological features considered to be typical of the senescent phenotype. The characteristic “flattening” of senescent cells was reflected by the decline in the density of staining (intensity of maximal pixel) of DNA-associated fluorescence [4,6-diamidino-2-phenylindole (DAPI)] paralleled by an increase in nuclear size (area). The decrease in ratio of maximal pixel to nuclear area was even more sensitive senescence biomarker than the change in maximal pixel or nuclear area, each alone. The saturation cell density at plateau phase of growth recorded by LSC was found to be dramatically decreased in cultures of senescent cells, thereby also serving as an additional marker. The induction of cyclin dependent kinase inhibitors p21(WAF1) and p27(KIP1) and γH2AX and activation of ATM markers of DNA damage response were measured in parallel with DNA/DAPI maximal pixel and nuclear area. These biomarker indices were expressed in quantitative terms by reporting them as a fraction of the respective controls. The effect of treatment of A549 and WI-38 cells with different concentrations of mitoxantrone (Mxt) and trichostatin A for various time periods was studied to assess the degree (depth) of cell senescence. Also assessed was the effect of 2-deoxy-d-glucose, the agent attenuating metabolic cell activity, on the depth of senescence induced by Mxt. A relationship between the ability of cells to synthesize RNA (incorporate 5-ethynyluridine) that leads to growth imbalance and induction of cell senescence was also studied. The data show that morphometric analysis of cellular attributes by LSC offers an attractive tool to detect cell senescence and measure its degree particularly in assessing effects of the factors that enhance or attenuate this process. This methodology is of importance in light of the evidence that cellular senescence is not only a biological process that is fundamental for organismal aging but also impedes formation of induced-pluripotent stem cells providing the barrier for neoplastic transformation and is the major mechanism of induction of reproductive cell death during treatment of solid tumors. © 2010 International Society for Advancement of Cytometry. Wiley Subscription Services, Inc., A Wiley Company 2010-11 2010-10-11 /pmc/articles/PMC2977923/ /pubmed/20939035 http://dx.doi.org/10.1002/cyto.a.20983 Text en Copyright © 2010 International Society for Advancement of Cytometry http://creativecommons.org/licenses/by/2.5/ Re-use of this article is permitted in accordance with the Creative Commons Deed, Attribution 2.5, which does not permit commercial exploitation.
spellingShingle Rapid Publication
Zhao, Hong
Halicka, H Dorota
Traganos, Frank
Jorgensen, Ellen
Darzynkiewicz, Zbigniew
New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry
title New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry
title_full New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry
title_fullStr New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry
title_full_unstemmed New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry
title_short New Biomarkers Probing Depth of Cell Senescence Assessed by Laser Scanning Cytometry
title_sort new biomarkers probing depth of cell senescence assessed by laser scanning cytometry
topic Rapid Publication
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC2977923/
https://www.ncbi.nlm.nih.gov/pubmed/20939035
http://dx.doi.org/10.1002/cyto.a.20983
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