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Immunofluorescence analysis of cytokeratin 8/18 staining is a sensitive assay for the detection of cell apoptosis
| Content Provider | Semantic Scholar |
|---|---|
| Author | Dong, Qiao-Mei Ling, Chun Zhao, Lu |
| Copyright Year | 2015 |
| 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. |
| Starting Page | 1227 |
| Ending Page | 1230 |
| Page Count | 4 |
| File Format | PDF HTM / HTML |
| Alternate Webpage(s) | https://www.spandidos-publications.com/ol/9/3/1227/download |
| PubMed reference number | 25663887 |
| Volume Number | 9 |
| Journal | Oncology letters |
| Language | English |
| Access Restriction | Open |
| Subject Keyword | Annexins Apoptosis Cell Death Cell Nucleus Chromosome Condensation Cisplatin Cytokeratin Cytoplasmic matrix DAPI Dyes Fluorescent Antibody Technique Immunostimulating conjugate (antigen) Iodides Isothiocyanate Microscopy, Fluorescence Neoplasms Phosphatidylserines Plasma membrane Propidium Iodide Staining method Tracer annexin A5 fluorescein sodium |
| Content Type | Text |
| Resource Type | Article |
