Visualization of mechanisms of kidney cancer A498 cell death under the influence of combined therapy of Gratīola officinālis extract and cyclophosphamide using the effect of fluorescence of ultrastructural cell components
Objective of the study: to evaluate the effectiveness of combined antitumor therapy with Gratīola officinālis extract in combination with synthetic cytostatic drug (cyclophosphamide). Materials and methods. Fluorescent dyes: iodine propidium and acridine orange. For imaging used a test "living and dead", which revealed the total number of dead cells by necrosis and apoptosis, the number of cells with apoptotic cells or nucleus pycnosis. The study was carried out within 24 hours, the dosage of drugs: Gratīola officinālis extract of 100 mcg/ml, cyclophosphamide 300 mcg/ml. The studied substances were used in 3 dosage ratios of Gratīola officinālis extract and cyclophosphamide (1:1; 1:2; 2:1), the comparison was made with the control group (without substances). Results. The most pronounced cytostatic and cytotoxic effect was observed at the ratio of the studied substances 1:1, with the total number of cells in the field of vision decreased by 4.5 times compared to the control group, the number of dead cells was increased by 3 times. In the study of the combination of Gratīola officinālis extract and cyclophosphamide in the ratio of dosages 1:2, a twofold decrease in the total number of cells in the field of vision was observed, but no pronounced cytotoxic effect was recorded. The ratio of dosages of the studied substances 2:1 showed a pronounced cytostatic effect (the total number of cells in the field of vision was 3 times less than in the control group). The highest apoptotic activity was in the dosage ratio of 1:1. Conclusions. The use of fluorescent dyes as a way to diagnose the mechanisms of cell death allowed studying the effect of combined antitumor therapy on cells of human kidney carcinoma A498. The most effective of the studied ratios of dosages of Gratīola officinālis extract and cyclophosphamide showed a ratio of 1:1, which combines a high level of cytostatic, cytotoxic, apoptotic activity.
A. M. Mylnikov
Saratov State Medical University n.a. V.I.Razumosky
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