Evaluation of human B cells response to exosomes secreted from breast cancer cells treated with cordycepin
Abstract
Breast cancer is one of the most important cancers based on its incidence and mortality rate, especially in women. One of the mechanisms of cancer progression is the inhibition of immune responses by cancer cells through the release of exosomes, which will be greater in metastatic conditions.
Methods: MDA-MB-231 cell line is cultured in RPMI1640 medium. One group of cells was treated with cordycepin and another solvent was added to the other group. After 24 hours, the supernatant is separated from the cells and their exosomes are purified using ultracentrifugation. Electron microscopy and DLS imaging will be used to confirm exosome extraction. In parallel, Raji cells are cultured and the exosomes extracted are affected in different amounts and the MTT method is examined for cell proliferation. The expression ratio of apoptotic and anti-apoptotic genes in these cells will be determined by real-time PCR. Flow cytometry is performed to determine the expression of CD19 marker
Results: Exosomes extracted from breast cancer cells treated with cordycepin decreased the proliferation of Raji cells, also decreased the expression of Bcl-2 gene and significantly increased the expression of BAX gene, and slightly increased the expression of CD19 marker.