Archives

  • 2022-07
  • 2022-06
  • 2022-05
  • 2022-04
  • 2021-03
  • 2020-08
  • 2020-07
  • 2020-03
  • 2019-11
  • 2019-10
  • 2019-09
  • 2019-08
  • 2019-07
  • br GW Inhibits the Release of BMSC

    2022-07-28


    GW4869 Inhibits the Release of BMSC-Derived Exosomes, Thus Suppressing miR-126-3p Expression
    In order to further investigate whether miR-126-3p carried by exosomes influences the biological functions of pancreatic cancer cells, the exo-somes’ secretory-specific inhibitor GW4869 was added to the co-culture system in the Transwell chamber to elucidate the effect of GW4869 on release of exosomes and the expression of miR-126-3p in pancreatic cancer cells. Compared with the NC DMSO group, the activity of acetyl-cholinesterase (AChE) in the GW4869 group was decreased, suggesting that the release of the exosomes and the miR-126-3p expression in the exosomes were reduced (p < 0.05; Figures 8A and 8B). Furthermore, the migration and invasion ability of pancreatic cancer Dorsomorphin was higher in the GW4869 group when compared with that of the DMSO group (p < 0.05; Figures 8C–8F). Based on the aforementioned results, it was concluded that GW4869 inhibited the release of BMSC-derived exosomes, resulting in the suppression of miR-126-3p expression.
    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 cells 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
    www.moleculartherapy.org
    A
    B
    C
    Proliferation rate (%)
    Number of migration cells
    Number of invasion cells 
    si-NC
    si-ADAM9
    si-NC
    si-ADAM9
    si-NC
    si-ADAM9
    si-NC
    Annexin V-FITC 
    Annexin V-FITC 
    Annexin V-FITC 
    Annexin V-FITC 
    si-NC
    E
    si-ADAM9
    F
    ADAM9
    GAPDH
    expression
    ADAM9
    Relative
    G
    si-NC
    si-ADAM9
    si
    expression 1.5
    si
    p inhibitor
    inhibit or
    protein
    Relative 0.5
    miR
    (legend on next page)
    Molecular Therapy: Nucleic Acids
    A C
    D
    Counts
    Counts
    Counts
    Counts
    Counts
    Counts
    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, 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