One of the tasks of current oncology is identification of cancer stem cells and search of therapeutic means capable of their specific inhibition. The paper presents the data on phenotype characteristics of Ehrlich carcinoma cells as convenient and easy-to-follow model of tumor growth. The evidence of cancer stem cells as a part of Ehrlich carcinoma and significance of CD44+ and CD44– subpopulations in maintaining the growth of this type of tumor were demonstrated. A high (tenfold) tumorigenic activity of the Ehrlich carcinoma CD44+ cells if compared to CD44– cells was proven. In this pair of comparison, the CD44+ cells had a higher potential of generating in peritoneal cavity of CD44high, CD44+CD24–, CD44+CD24+ cell subpopulations, highlighting the presence of cancer stem cells in a pool of CD44+ cells.In this study, the ability of synthesized hybrid nanocomplexes, comprising the nanoparticles of rare earth orthovanadates GdYVO4:Eu3+ and cholesterol to inhibit the tumor growth and to increase the survival of the animals with tumors was established. A special contribution into tumor-inhibiting effect is made by each of its components. Treatment of Ehrlich carcinoma cells with two-component hybrid complex resulted in maximum reduction in the concentration of the most tumorigenic CD44high cells with simultaneous rise in the number of CD117+ cells that decreased an intensity of tumor growth by 74.70 ± 4.38% if compared with the control.
У роботі представлений метод виділення мононуклеарів з кісткового мозку у градієнті «тразографу», який дає змогу отримати більш ніж 40 % клітин, серед яких 28 %-CD14 +-моноцити. Підібрана мінімальна панель фенотипічних маркерів для оцінки незрілих дендритних клітин у культурі in vitro. В порівняльному аспекті продемонстровано, що більш адекватним джерелом отримання дендритних клітин є мононуклеари на відміну від загальної суспензії клітин кісткового мозку. Встановлено, що диференціювання мієлоїдних попередників і моноцитів у незрілі дендритні клітини супроводжується втратою CD14-антигена, мінімізацією рівня експресії костимуляторної молекули СD86 і появою характерного для цих клітин маркера CD11b-антигена. Результати дослідження дають змогу не тільки оцінити якісні характеристики отриманих дендритних клітин і їх толерогенну активність на моделі аутоімунних захворювань в експерименті, а також намітити перспективи створення їх запасів у низькотемпературних банках для клінічного використання.
A most important task in oncology today is the search for structures capable of specific targeting and inactivating the cancer stem cells determining the tumor growth. We describe the investigation of the ability of the synthesized hybrid nanocomplexes containing rare earth-based nanoparticles of GdYVO 4 : Eu 3+ orthovanadates, cholesterol and the hydrophobic luminescent dye DiI to visualize and inactivate the cancer stem cells of the Ehrlich carcinoma. The spectral analysis revealed an intracellular accumulation of nanoparticles in the Ehrlich carcinoma cells. This fact suggested the implementation of antitumor effect of the nanoparticles in vivo. The treatment of the Ehrlich carcinoma cells with spherical nanoparticles at a concentration of 1.30 g/L was found to reduce the intensity of tumor growth by 58.2 %. Ehrlich carcinoma cell processing with a three component hybrid complex resulted in a maximum decrease in the concentration of the most carcinogenic CD44 hi cells with a simultaneous increase in the number of CD117 + cells, which reduced the intensity of tumor growth by 76.6 %. When applying hybrid nanocomplexes, a reduced expression rate of the oct4, sox2 and nanog genes in Ehrlich carcinoma cells was observed which may be a mechanism for the antitumor action of orthovanadates.
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