THE EFFECT OF DOCETAXEL ON THE SURVIVAL AND CLONOGENIC ACTIVITY OF MCF-7 MAMMOSPHERES CULTURE ENRICHED WITH CANCER STEM CELLS

DOI: https://doi.org/10.29296/24999490-2019-05-09

V.G. Shuvatova, Yu.P. Semochkina, G.A. Posypanova, E.Yu. Moskaleva National Research Center «Kurchatov Institute», Akademika Kurchatova square, 1, Moscow, 123182, Russian Federation E-mail: [email protected]

Introduction. Cancer stem cells (CSCs), which are less sensitive to chemotherapeutic drugs than other tumor cells, play a key role in the initiation and development of the tumor process. Maintenance of CSCs determines the risk of cancer recurrence. The aim of the study. To study the effect of docetaxel on CSCs survival and self-renewal in the human mammary adenocarcinoma MCF-7 cells cultured as mammospheres. Methods. The effect of docetaxel on the CSCs survival was studied after 72 and 120 h of cultivation with the drug at concentrations of 0, 1, 10 and 100 nmol/l. For this purpose, the total number of living cells was counted, the fraction of CSCs with CD44+/CD24-/low phenotype was determined using flow cytometry, and the absolute number of CSCs was calculated in each sample. The effect of docetaxel on the CSCs repopulation was determined by counting mammospheres formed 7 days after cells were washed from docetaxel after 120 h cultivation with the drug at concentrations 0, 1, 10 and 100 nmol/l. Results. The cultivation of mammospheres with docetaxel for 72 h resulted in an insignificant decrease in both of the total cell number and the CSCs fraction. At the same time, there was a significant decrease in the number of CSCs. The cultivation of mammospheres at all studied docetaxel concentrations for 120 hours resulted in a significant decrease in the total number of cells, the proportion and the number of CSCs and their clonogenic activity that represents the ability of the CSC to repopulate. Conclusion. CSCs of MCF-7 mammospheres were shown to be highly sensitive to docetaxel after prolonged cultivation (120 h). Docetaxel inhibited the ability of CSCs to self-renewal, which plays a crucial role in the tumor and metastasis initiation
Keywords: 
docetaxel, cancer stem cells, clonogenic activity, human breast adenocarcinoma, MCF-7, mammospheres
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