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  • br miR p Transferred by BMSC Derived Exosomes

    2020-08-18


    miR-126-3p Transferred by BMSC-Derived Exosomes Suppresses Proliferation, Migration, and Invasion of Pancreatic Cancer Cells and Enhances Their Apoptosis
    In order to examine the effect of miR-126-3p in BMSC-derived exo-somes on the biological function of pancreatic cancer cells, the cell 
    proliferation, migration, and invasion abilities of pancreatic cancer LDN-193189 were examined in BMSC-derived exosomes overexpressing miR-126-3p. The results obtained indicated that (Figures 9A–9D), in comparison with the BMSCs-miR-126-3p NC group, the cell pro-liferation, migration, and invasion abilities of the BMSCs-miR-126-3p mimic group were markedly elevated, whereas the apoptosis rate was notably increased (p < 0.05). Western blot analysis revealed that the expression of proliferation-related factors [Ki67 and vascular endothelial growth factor (VEGF)] and invasive factors [cyclooxyge-nase-2 (COX-2) and matrix metalloproteinase-14 (MMP-14)] in the BMSCs-miR-126-3p mimic group was significantly lower than that in the BMSCs-miR-126-3p NC group (all p < 0.05; Figures 9E and 9F). The obtained data suggested that BMSCs deliver miR-126-3p via exo-somes to inhibit proliferation, migration, and invasion of pancreatic cancer cells and promote their apoptosis.
    miR-126-3p in BMSC-Derived Exosomes Inhibits Tumor Growth and Metastasis of Pancreatic Cancer Cells
    In order to investigate the effect of miR-126-3p on tumor growth, overexpressed miR-126-3p in BMSCs was examined by xenograft in nude mice. In contrast with the BMSCs-miR-126-3p NC group, the
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    Figure 6. Results of BMSCs Sorting
    (A) Morphology of BMSCs in primary adherent culture (original magnification 100). (B) The morphology of the third generation BMSCs observed under a microscope (original magnification 200). (C) Positive markers (CD29, CD44, and CD71) and negative markers (HLA-DR, LDN-193189 CD34, and CD45) of BMSCs identified by flow cytometry. (D) Results of oil red O staining after 2 weeks of lipid-induced differentiation (original magnification 400). (E) Results of alizarin red and Von Kossa staining after 4 weeks of osteogenic induction (original magnification 400). The data were all measured and expressed by mean ± SD. The independent sample t test was used for statistical analysis between two groups, and the experiment was repeated three times. BMSC, human bone mesenchymal stem cell.
    tumor formation time was shorter, along with a slower growth rate, as well as smaller tumors, in the BMSCs-miR-126-3p mimic group (p < 0.05; Figures 10A–10C). The expression of miR-126-3p and ADAM9 was verified through the application of qRT-PCR and west-ern blot analysis methods, respectively. Compared with the BMSCs-miR-126-3p NC group, the expression of miR-126-3p in the BMSCs-miR-126-3p mimic group was increased significantly (p < 0.05; Figure 10D), whereas that of ADAM9 decreased in a dra-matic fashion (p < 0.05; Figure 10D–10F). The expression of COX-2 and MMP-14 in the BMSCs-miR-126-3p mimic group was decreased when compared with the BMSCs-miR-126-3p NC group (p < 0.05; Figure 10D). The results obtained demonstrated that the miR-126-3p expression in the BMSCs-miR-126-3p mimic group was markedly elevated (p < 0.05; Figures 10E and 10F). Taken together, the results obtained suggested that BMSC-derived exosomes transferred miR-126-3p to inhibit proliferation, invasion, and metas-tasis of pancreatic cancer cells.
    DISCUSSION
    As the fourth main cause of cancer-related mortality, pancreatic can-cer has had a survival rate of less than 5% for many decades due to few effective therapies.3 miRNAs have been highlighted because of their
    potential as cancer biomarkers and close association with the pro-cesses of tumorigenesis and various physiological and pathological
    developments including cell proliferation, differentiation, and apoptosis.16,22 In our study, the specific mechanism by which miR-
    126-3p transferred via BMSC-derived exosomes influenced the bio-logical function of pancreatic cancer was identified. The findings demonstrated that miR-126-3p shuttled by BMSC-derived exosome, by downregulating ADAM9, suppressed proliferation, invasion, and metastasis and promoted apoptosis in pancreatic cancer cells, thus acting as a potential biomarker for the treatment of pancreatic cancer.