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    concentration (particies/ml)
    control -exo
    level in BMSCs 
    level in Exosome 
    F ADAM9
    expression of
    NC mimic
    ADAM9 protein Relative expression 
    Figure 7. miR-126-3p Could Be Carried by BMSC-Derived Exosomes
    (A) The ultrastructure of exosomes observed by TEM (original magnification 400). (B) Size distribution of exosomes detected by NanoSight particle size analysis. (C) Surface markers of exosomes (CD63, Hsp70) detected by western blot analysis. (D) The exosomes absorbed by PANC-1 Pyocyanin under confocal fluorescence microscope at different time points. Red: miR-126-Cy3; green: pCDNA3.1-GFP; blue; DAPI. (E) The expression of miR-126-3p in BMSCs, exosomes, and co-cultured pancreatic cancer cells determined by qRT-PCR. (F) mRNA expression of ADAM9 in pancreatic cancer cells determined by qRT-PCR. (G and H) Protein expression of ADAM9 in pancreatic cancer cells determined by western blot analysis. The data in the chart were analyzed using an independent sample t test, and the experiment was repeated three times. *p < 0.5. BMSC, bone marrow mesenchymal stem cell; GAPDH, glyceraldehyde-3-phosphate dehydrogenase; miR-126-3p, microRNA-126-3p; NC, negative control; TEM, transmission electron microscopy.
    Molecular Therapy: Nucleic Acids
    Relative expression 
    migration cells
    cell number
    Figure 8. GW4869 Inhibited the Release of BMSC-Derived Exosomes to Inhibit the Expression of miR-126-3p
    (A) The effect of GW4869 on the release of exosomes detected by AChE activity assay. (B) The effect of GW4869 on the expression of miR-126-3p in pancreatic cancer cells determined using qRT-PCR. (C and E) The effect of GW4869 on migration ability of PANC-1 detected by Transwell assay (photograph of migration cell, C, and number of migration cell, E). (D and F) The effect of GW4869 on invasion of PANC-1 determined by Transwell assay (photograph of invasion cell, D, and number of migration cell, F). The data were measured by the mean ± SD. Comparison between two groups was conducted using an independent sample t test for statistical analysis; *p < 0.05, compared with the DMSO group. The experiment was repeated three times. miR-126-3p, microRNA-126-3p.
    Existing literature has suggested that ADAM9 is overexpressed dur-ing the progression of cancer in addition to indicating that silencing ADAM9 could promote both the radio-sensitivity and chemosensi-tivity of cancer cells to therapeutic drugs.33 Statistical evidence has demonstrated that pancreatic cancer cells exhibit a significantly higher level of ADAM9, which was also observed in the current study.34 Liu et al.20 concluded that miR-126 could suppress the expression of ADAM9 and act to further inhibit the growth of the ESCC through the epidermal growth factor receptor (EGFR)-AKT pathway. Various reports have pointed to the fact that pancreatic can-cer cells exhibit downregulated levels of miR-126-3p and upregulated ADAM9, in addition to revealing that ADAM9 is a target gene of miR-126-3p, which acts to directly inhibit ADAM9.
    Exosomes are membrane-enclosed small vesicles (30–100 nm) with endosomal origin that contain numerous molecular components including proteins, mRNAs, as well as miRNAs.35 Exosomes secreted by cells play a crucial role in correlating cells and transmit-ting information among tissues over long distances.36 BMSCs have been demonstrated to interact with tumor cells and participate in the progression of multiple tumors, including the processes of cell migration, invasion, and proliferation.37 More specifically, miR-31a-5p transferred by BMSC-derived exosomes has been shown to influence osteogenesis and osteoclastic differentiation.38 During the current study, a key observation was made indicating that BMSC-derived exosomes enriched miR-126-3p. Additionally, miR-126 enriched in exosomes from normal endothelial cells has been reported to contribute to suppressing cell growth and reducing the malignancy of non-small cell lung cancer cells.39 Studies have suggested that exosomes derived from BMSCs could alleviate liver injury of autoimmune hepatitis, which is correlated to the negative regulation of exosomal miR-233 on nucleotide-binding