Flow cytometric analysis of apoptotic biomarkers in actinomycin D-treated SiHa cervical cancer cells

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dc.contributor.author Punchoo, Rivak
dc.contributor.author Zhou, Esther
dc.contributor.author Bhoora, Sachin
dc.date.accessioned 2022-09-14T06:41:07Z
dc.date.available 2022-09-14T06:41:07Z
dc.date.issued 2021-08-26
dc.description.abstract Apoptosis biomarkers were investigated in actinomycin D-treated SiHa cervical cancer cells using a benchtop flow cytometer. Early biomarkers (Annexin V and mitochondrial membrane potential) and late biomarkers (caspases 3 and 7, and DNA damage) of apoptosis were measured in experimental and control cultures. Cultures were incubated for 24 hours in a humidified incubator at 37 °C with 5% CO2. The cells were then detached using trypsin and enumerated using a flow cytometric cell count assay. Cells were further analyzed for apoptosis using an Annexin V assay, a mitochondrial electrochemical transmembrane potential assay, a caspase 3/7 assay, and a DNA damage assay. This article provides an overview of apoptosis and traditional flow cytometry, and elaborates flow cytometric protocols for processing and analyzing SiHa cells. The results describe positive, negative, and sub-optimal experimental data. Also discussed are interpretation and caveats in performing flow cytometric analysis of apoptosis using this analytical platform. Flow cytometric analysis provides an accurate measurement of early and late biomarkers for apoptosis. en_US
dc.description.department Chemical Pathology en_US
dc.description.librarian am2022 en_US
dc.description.sponsorship The National Research Foundation (NRF) and the South African Medical Research Council (SAMRC). en_US
dc.description.uri https://www.jove.com/journal en_US
dc.identifier.citation Punchoo, R., Zhou, E., Bhoora, S. Flow Cytometric Analysis of Apoptotic Biomarkers in Actinomycin D-treated SiHa Cervical Cancer Cells. Journal of Visualized Experiments (174), e62663, DOI:10.3791/62663 (2021). en_US
dc.identifier.issn 1940-087X (online)
dc.identifier.other 10.3791/62663
dc.identifier.uri https://repository.up.ac.za/handle/2263/87176
dc.language.iso en en_US
dc.publisher MyJove Corporation en_US
dc.rights © 2021 JoVE Creative Commons Attribution-NonCommercial-NoDerivs 3.0 Unported License. en_US
dc.subject Apoptosis biomarkers en_US
dc.subject Flow cytometer en_US
dc.subject Deoxyribonucleic acid (DNA) en_US
dc.subject D-treated SiHa cervical cancer cells en_US
dc.subject Cervical cancer en_US
dc.subject.other Health sciences articles SDG-03
dc.subject.other SDG-03: Good health and well-being
dc.title Flow cytometric analysis of apoptotic biomarkers in actinomycin D-treated SiHa cervical cancer cells en_US
dc.type Article en_US


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