Optimized fluorescent probe combinations for evaluation of proliferation and necrosis in anthracycline‐treated leukaemic cell lines
| Autor: | M. Mousseau, M. Barbier, D. Seigneurin, Jean Boutonnat, Xavier Ronot, K. A. Muirhead |
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| Jazyk: | angličtina |
| Rok vydání: | 2003 |
| Předmět: |
Necrosis
Indoles Cell division Daunorubicin Cytological Techniques Color Indicator Dilution Techniques Biology Flow cytometry chemistry.chemical_compound Organophosphorus Compounds medicine Organometallic Compounds Humans Scattering Radiation Propidium iodide DAPI Coloring Agents health care economics and organizations Fluorescent Dyes Antibiotics Antineoplastic medicine.diagnostic_test Cell growth Cell Membrane Cell Biology General Medicine Original Articles Molecular biology Antineoplastic Agents Phytogenic humanities chemistry Biochemistry Microscopy Fluorescence Cell culture Vincristine medicine.symptom K562 Cells Biomarkers Cell Division medicine.drug Propidium |
| Popis: | Proliferation and multidrug resistance status are key predictors of therapeutic outcome in acute myeloid leukaemia (AML). Anthracyclines such as daunorubicin (DNR) are typically used to treat AML and can induce drug resistance. The goal of the studies described here was to select a combination of fluorescent probes that could be used in combination with flow cytometry to monitor cell proliferation vs. cell death/necrosis as a function of anthracycline uptake. Propidium iodide (PI), the most commonly used marker of membrane integrity, cannot be used to evaluate necrosis in DNR‐containing cells because of spectral overlap. A membrane integrity probe compatible with the use of a dye dilution method using PKH67 to study cell proliferation was also selected. The results show that DAPI and Cascade Blue (CB), like PI, were able to detect necrotic cells when no DNR was present, although CB gave less resolution between viable and necrotic cells than PI or DAPI. In the presence of DNR, DAPI cannot be used owing to the fluorescence quenching by DNR. However, it was found that a combination of DNR, CB, and PKH67 allows simultaneous identification of chemoresistant cells, based on reduced DNR accumulation, necrotic cells based on CB incorporation, and proliferating cells based on partitioning of PKH67 fluorescence between daughter cells. It was also found that unless a marker of necrosis is used in combination with the dye dilution assay, a moderate decrease of fluorescence as a result of necrosis may be incorrectly interpreted as proliferation. |
| Databáze: | OpenAIRE |
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