Abstract
Abstract Exosome is referred as membrane-bound nano-vesicles secreted by various mammalian cells. It can be found in different body fluids, such as plasma, saliva, urine, latex, cerebrospinal fluid, ascites, and cell supernatant, which contains a large number of proteins, miRNAs, mRNAs, lipids, nucleic acids, peptides and other substances. Exosome plays a pivotal role in the information transmission signals between cells and the diagnosis and prognosis of diseases as well as drug evaluation. It has become an emerging field in cancer research recently. Therefore, we separated exosomes from human kidney embryo cell (HEK-293) by ultra high speed centrifugation. To identify exosomes from their surface specific proteins by Western blotting analysis. We found enrichment of exosome markers CD63 and TSG101 without Calnexin protein in the exosoms so confirmed that exosome was successfully isolated. Then, we transfered mRNA let-7b into the exosomes by electric transfection further evaluated the effects of cytotoxicity and apoptosis induction on human breast cancer MCF-7 cells by CCK 8 assay Hochest 33342-PI and AnnexinV-PI dual staining analysis, respectively. The results showed that exosome mediated microRNA let-7b exhibited significant anticancer activity and apoptosis induction in a concentration-dependent manner in MCF-7 cells. Moreover, exosome mediated let-7b significantly up-regulated the expression levels of apoptosis related proteins casepase 3, casepase 9, and PARP determined by Western blotting analysis. Our results indicate that the exosomes mediated microRNA such as let-7b can effectively inhibit cell growth and induce apoptosis against breast cancer cells and it may be developed as a novel anticancer agent in the treatment of cancer clinically. Citation Format: Xu Biantiao. Anticancer effect and mechanism of exosomes mediated microRNA let-7b in breast cancer cells [abstract]. In: Proceedings of the American Association for Cancer Research Annual Meeting 2019; 2019 Mar 29-Apr 3; Atlanta, GA. Philadelphia (PA): AACR; Cancer Res 2019;79(13 Suppl):Abstract nr 725.
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