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Published on: February 16, 2015
Ethanol induced apoptosis in human HL-60 cells
M Mareková1, J Vávrová, D Vokurková
1Department of Medical Biochemistry, Faculty of Medicine Hradec Králové, Charles University Prague, Czech Republic. marekova@lfhk.cuni.cz
General Physiology and Biophysics
|January 13, 2001
Summary
Morphological changes and flow cytometry detect apoptosis in HL-60 cells early, while viability assays show changes later. Ethanol-induced apoptosis in these leukemia cells reveals distinct cellular events over time.
Area of Science:
- Cell Biology
- Hematology
- Apoptosis Research
Background:
- Apoptosis detection methods are crucial for understanding cell death in diseases like leukemia.
- Human promyelocytic leukemia cell line HL-60 is a standard model for studying apoptosis.
Purpose of the Study:
- To compare the efficacy of flow cytometry and morphologic methods in detecting apoptosis in HL-60 cells.
- To analyze the temporal dynamics of apoptosis induction by ethanol in HL-60 cells.
Main Methods:
- Induction of apoptosis in HL-60 cells using 3% ethanol.
- Assessment of apoptosis via flow cytometry (forward scatter, APO2.7 antibody staining, sub-diploid DNA content) and light microscopy (Diff-Quik staining).
- Viability assessment using Trypan-blue staining and cell surface marker analysis (CD95).
Main Results:
- Morphological changes and sub-G1 DNA content indicative of apoptosis were detected by 4 hours, preceding significant changes in forward scatter or APO2.7 staining.
- Flow cytometry and morphology showed nearly all cells undergoing apoptosis by 7 hours.
- Cell size (forward scatter) decreased significantly by 16 hours, with increased APO2.7 positivity.
- By 24 hours, only a small fraction of viable, APO2.7-negative cells remained; all cells were non-viable by 48 hours.
- Ethanol treatment did not increase CD95 expression on HL-60 cells.
Conclusions:
- Morphological and flow cytometric methods, particularly sub-diploid DNA content and APO2.7 staining, are sensitive early indicators of ethanol-induced apoptosis in HL-60 cells.
- Viability assays and forward scatter changes reflect later stages of apoptosis.
- HL-60 cells are characterized as CD34-/AC133-, CD33+/CD15+, with low CD95 expression, and this expression is not upregulated during ethanol-induced apoptosis.
