Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis

  • Authors:
    • Qiao‑Mei Dong
    • Chun Ling
    • Li Zhao
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  • Published online on: January 7, 2015     https://doi.org/10.3892/ol.2015.2856
  • Pages: 1227-1230
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Abstract

Apoptosis is one  of the major types of programmed cell death. During this process, cells experience a series of morphological and biochemical changes. Flow cytometric analysis of Annexin V staining of cell surface phosphatidylserine, in combination with a DNA‑staining dye to probe the permeability of the cell membrane, is an established method for detecting apoptosis. The present study aimed to show that the immunofluorescence analysis of cytokeratin (CK) 8/18 staining may provide a new and sensitive assay for the detection of apoptotic cells. Tumor cells were treated with 20 µM cisplatin to induce apoptosis. Following 12 and 24 h of cisplatin treatment, cells were collected and stained with 4',6‑diamidine‑2'‑phenylindole dihydrochloride (DAPI) and fluorescein‑labeled anti‑CK8/18 antibody. The apoptotic cells were subsequently examined by fluorescence microscopy. Annexin V‑fluorescein isothiocyanate/propidium iodide staining followed by flow cytometric analysis confirmed that cisplatin was able to induce apoptosis in tumor cells. Immunofluorescence analysis demonstrated that apoptotic cells had a distinct CK8/18 staining pattern. In living cells, CK8/18 was uniformly distributed in the cytoplasm and cytosol; however in the apoptotic cells with a condensed and/or fragmented apoptotic nucleus (as identified by DAPI staining), fluorescein‑labeled anti‑CK8/18 antibody exhibited unusual punctate and/or bubbly staining in the cytosol. In the apoptotic cells that could not be identified by DAPI staining, fluorescein‑labeled CK8/18 displayed polarized aggregated staining in the cytosol. These results indicate that fluorescein‑conjugated CK8/18 may be a useful and sensitive indicator of cell apoptosis.
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March-2015
Volume 9 Issue 3

Print ISSN: 1792-1074
Online ISSN:1792-1082

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Spandidos Publications style
Dong QM, Ling C and Zhao L: Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis. Oncol Lett 9: 1227-1230, 2015.
APA
Dong, Q., Ling, C., & Zhao, L. (2015). Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis. Oncology Letters, 9, 1227-1230. https://doi.org/10.3892/ol.2015.2856
MLA
Dong, Q., Ling, C., Zhao, L."Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis". Oncology Letters 9.3 (2015): 1227-1230.
Chicago
Dong, Q., Ling, C., Zhao, L."Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis". Oncology Letters 9, no. 3 (2015): 1227-1230. https://doi.org/10.3892/ol.2015.2856