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Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-Treated SiHa Cervical Cancer Cells
Published on: August 26, 2021
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Flow cytometry-based quantitative analysis of cellular protein expression in apoptosis subpopulations: A protocol
Salah Abdalrazak Alshehade1, Hassan A Almoustafa2,3, Mohammed Abdullah Alshawsh2,4
1Department of Pharmacology, Faculty of Pharmacy & Bio Medical Sciences, MAHSA University, 42610, Selangor, Malaysia.
Heliyon
|July 23, 2024
Summary
This study presents a flow cytometry protocol to track apoptosis and protein changes in cancer cells. It enables quantitative analysis of cell death and CD44 expression during doxorubicin treatment.
Area of Science:
- Cell Biology
- Immunology
- Biochemistry
Background:
- Apoptosis is a critical cellular process.
- Flow cytometry is a powerful tool for cell analysis.
- Quantitative apoptosis assessment requires precise methodologies.
Purpose of the Study:
- To develop a multiparametric flow cytometry protocol for apoptosis analysis.
- To simultaneously track protein expression changes during apoptosis.
- To investigate doxorubicin-induced apoptosis and CD44 expression in breast cancer cells.
Main Methods:
- Dual staining with annexin V-FITC and propidium iodide (PI) for apoptosis detection.
- Conjugation of APC-labeled antibodies for simultaneous protein analysis.
- Application in MDA-MB-231 breast cancer cells treated with doxorubicin.
Main Results:
- Quantitative assessment of viable, early apoptotic, and late apoptotic/necrotic cell populations.
- Demonstration of decreased CD44 expression correlating with apoptosis progression.
- Successful integration of apoptosis and protein expression analysis.
Conclusions:
- The developed protocol provides robust quantitative insights into apoptosis.
- It facilitates the study of signaling networks and therapeutic resistance.
- This method is applicable to various cellular models for apoptosis research.
