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    A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue

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    Author
    Wang, Mengdie
    Knudsen, Beatrice S
    Nagle, Raymond B
    Rogers, Gregory C
    Cress, Anne E
    Affiliation
    Univ Arizona, Canc Biol Res Program
    Univ Arizona, Dept Pathol
    Univ Arizona, Dept Cellular & Mol Med
    Univ Arizona, Univ Arizona Canc Ctr
    Issue Date
    2019-03-21
    
    Metadata
    Show full item record
    Publisher
    AMER SOC CELL BIOLOGY
    Citation
    Wang, M., Knudsen, B. S., Nagle, R. B., Rogers, G. C., & Cress, A. E. (2019). A method of quantifying centrosomes at the single-cell level in human normal and cancer tissue. Molecular biology of the cell, 30(7), 811-819.
    Journal
    MOLECULAR BIOLOGY OF THE CELL
    Rights
    © 2019 Wang et al. This article is distributed by The American Society for Cell Biology under license from the author(s). Two months after publication it is available to the public under an Attribution–Noncommercial–Share Alike 3.0 Unported Creative Commons License.
    Collection Information
    This item from the UA Faculty Publications collection is made available by the University of Arizona with support from the University of Arizona Libraries. If you have questions, please contact us at repository@u.library.arizona.edu.
    Abstract
    Centrosome abnormalities are emerging hallmarks of cancer. The overproduction of centrosomes (known as centrosome amplification) has been reported in a variety of cancers and is currently being explored as a promising target for therapy. However, to understand different types of centrosome abnormalities and their impact on centrosome function during tumor progression, as well as to identify tumor subtypes that would respond to the targeting of a centrosome abnormality, a reliable method for accurately quantifying centrosomes in human tissue samples is needed. Here, we established a method of quantifying centrosomes at a single-cell level in different types of human tissue samples. We tested multiple anti-centriole and pericentriolar-material antibodies to identify bona fide centrosomes and multiplexed these with cell border markers to identify individual cells within the tissue. High-resolution microscopy was used to generate multiple Z-section images, allowing us to acquire whole cell volumes in which to scan for centrosomes. The normal cells within the tissue serve as internal positive controls. Our method provides a simple, accurate way to distinguish alterations in centrosome numbers at the level of single cells.
    ISSN
    1939-4586
    PubMed ID
    30699045
    DOI
    10.1091/mbc.E18-10-0651
    Version
    Final published version
    Sponsors
    National Cancer Institute (NCI) [P30CA23074]; Department of Defense Prostate Cancer Research Program [W81XWH-14-2-0182, W81XWH-14-2-0183, W81XWH-14-2-0185, W81XWH-14-2-0186, W81XWH-15-2-0062]; National Institutes of Health (NIH) [R01GM110166, R01GM126035]; NIH-NCI [RO1CA159406]; Tim and Diane Bowden Cancer Biology Research Fellowship
    Additional Links
    https://www.molbiolcell.org/doi/full/10.1091/mbc.E18-10-0651
    ae974a485f413a2113503eed53cd6c53
    10.1091/mbc.E18-10-0651
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