Fluorescent Methods for the Visualization of Apoptosis and Autophagy in Human Tumor Cells
Nikita A. Navolokin, 1,2, Natalya V. Polukonova,1, Dmitriy A. Mudrak,1, Artyem M. Myl’nikov,1, Mariya A. Baryshnikova,3, Dmitriy A. Khochenkov,3, Alla B. Bucharskaya,1,2, Anna V. Polukonova,1, Galina N. Maslyakova,1,2, 1 Saratov State Medical University n.a.V.I.Razumovsky, Saratov, 2 Saratov State University, Saratov, Russia, 3 Blokhin Center for Medical Oncology Research, Moscow, Russia
Abstract
The potential use of fluorescent methods and their advantages for visualizing and identification of the type of programmed cell death was examined in human tumor cells exposed to flavonoids. HeLa cervical cancer cells and A498 renal cell carcinoma cells were exposure the flavonoid-containing extract of Gratiola officinalis L. as an experimental treatment. The following fluorescent methods were used: “live” and “dead” test with double staining with propidium iodide and acridine orange. Induction of autophagy was confirmed by fluorescent tests performed on a Muse Cell Analyzer using the Muse Autophagy LC3-Antibody Based Kit. The double staining with acridine orange and propidium iodide fluorescent dyes in the live and dead test and comparison with phase-contrast microscopy allows to visualize the formation processes of apoptotic bodies and autophagosomes, therefore, can be used as a method of screening evaluation of the effectiveness of various types of treatment.
Speaker
Nikita Navolokin
Saratov State Medical University n.a.V.I.Razumovsky
Russia
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