Research Paper Volume 13, Issue 4 pp 5055—5068

MicroRNA-29a-3p delivery via exosomes derived from engineered human mesenchymal stem cells exerts tumour suppressive effects by inhibiting migration and vasculogenic mimicry in glioma

Transfected MSCs transferred miR-29a-3p via exosomes and inhibited migration and VM formation in glioma. (A) Representative images of human MSCs transfected with miR-29a-3p or an NC nucleotide sequence (scale bar, 100 μm) and the corresponding electron microscopic images of exosomes (scale bar=100 nm). (B) Nanoparticle tracking technology indicated an accumulation of particles of diameters of 50-100 nanometres. (C) Western blot analysis showing the presence of TSG101 and CD9 and the absence of calnexin in MSC-derived exosomes. Results are from three independent experiments. (D, E) PCR analysis of the miR-29a-3p level in exosomes (D) and glioma cell lines pretreated with exosomes (E). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P F) Migration capability detected by a transwell assay after treatment with miR-29a-3p-overexpressing exosomes (scale bar, 100 μm; n=3). (G) Quantification of transwell migration assays in (F). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P H) VM formation after treatment with miR-29a-3p-overexpressing exosomes (scale bar, 200 μm; n=3). (I) Quantification of relative VM numbers in (H). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P J) Protein levels of markers of migration and VM formation detected by western blotting after miR-29a-3p-overexpressing exosome (EXO-29a) treatment. Results are from three independent experiments.

Figure 4. Transfected MSCs transferred miR-29a-3p via exosomes and inhibited migration and VM formation in glioma. (A) Representative images of human MSCs transfected with miR-29a-3p or an NC nucleotide sequence (scale bar, 100 μm) and the corresponding electron microscopic images of exosomes (scale bar=100 nm). (B) Nanoparticle tracking technology indicated an accumulation of particles of diameters of 50-100 nanometres. (C) Western blot analysis showing the presence of TSG101 and CD9 and the absence of calnexin in MSC-derived exosomes. Results are from three independent experiments. (D, E) PCR analysis of the miR-29a-3p level in exosomes (D) and glioma cell lines pretreated with exosomes (E). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P < 0.05). (F) Migration capability detected by a transwell assay after treatment with miR-29a-3p-overexpressing exosomes (scale bar, 100 μm; n=3). (G) Quantification of transwell migration assays in (F). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P < 0.05). (H) VM formation after treatment with miR-29a-3p-overexpressing exosomes (scale bar, 200 μm; n=3). (I) Quantification of relative VM numbers in (H). Data are shown as the mean±SD, n=3, one-way ANOVA (*, P < 0.05). (J) Protein levels of markers of migration and VM formation detected by western blotting after miR-29a-3p-overexpressing exosome (EXO-29a) treatment. Results are from three independent experiments.