09 February 2015 : Original article
Med Sci Monit Basic Res 2015; 21:15-20
BACKGROUND: The aim of this study was to evaluate the ability of dual acridine orange/ethidium bromide (AO/EB) staining to detect tumor cell apoptosis. According to apoptosis-associated changes of cell membranes during the process of apoptosis, a clear distinction is made between normal cells, early and late apoptotic cells, and necrotic cells.
MATERIAL AND METHODS: We cultured human osteosarcoma cells with 30, 60, and 120 µg/ml kappa-selenocarrageenan. To assess the rates of cell proliferation and apoptosis, cells were fluorescently stained with acridine orange/ethidium bromide (AO/EB) or stained with propidium iodide (PI) and analyzed by flow cytometry. All experiments were repeated at least 3 times.
RESULTS: Normal tumor cells, early and late apoptotic cells, and necrotic cells were examined using fluorescent microscopy. Early-stage apoptotic cells were marked by crescent-shaped or granular yellow-green acridine orange nuclear staining. Late-stage apoptotic cells were marked with concentrated and asymmetrically localized orange nuclear ethidium bromide staining. Necrotic cells increased in volume and showed uneven orange-red fluorescence at their periphery. Cells appeared to be in the process of disintegrating. The percentage of apoptotic osteosarcoma cells detected by dual acridine orange/ethidium bromide (AO/EB) staining was not significantly different from that detected using flow cytometry (P>0.05).
CONCLUSIONS: Our results suggest that dual acridine orange/ethidium bromide staining is an economic and convenient method to detect apoptosis in tumor cells and to test tumor chemosensitivity compared with flow cytometry.
Keywords: Apoptosis - physiology, Acridine Orange - diagnostic use, Carrageenan, Cell Proliferation - physiology, Ethidium - diagnostic use, Organoselenium Compounds, Osteosarcoma - physiopathology, Propidium, Staining and Labeling - methods
Most Viewed Current Articles
05 Jan 2021 : Review articleA Southeast Asian Perspective on the COVID-19 Pandemic: Hemoglobin E (HbE)-Trait Confers Resistance Against...
Med Sci Monit Basic Res 2021; 27:e929207
05 May 2022 : Laboratory ResearchCalcitriol Inhibits Proliferation and Potentially Induces Apoptosis in B16-F10 Cells
Med Sci Monit Basic Res 2022; 28:e935139
09 Jun 2021 : Laboratory ResearchVitamin D Inhibits Lipopolysaccharide (LPS)-Induced Inflammation in A549 Cells by Downregulating Inflammato...
Med Sci Monit Basic Res 2021; 27:e931481
07 Jul 2022 : Laboratory ResearchCytotoxicity, Apoptosis, Migration Inhibition, and Autophagy-Induced by Crude Ricin from Ricinus communis S...
Med Sci Monit Basic Res 2022; 28:e936683