Conclusions
Our data indicated that exosomes can be suitable carriers for miR-211 mimic. Moreover, TEXomiR via anti-cancer effects could inhibit the progression of melanoma cancer.
Material and methods
After exposing B16F10 cultured cells to serum-free media for 24 hours, we collected the supernatant. Subsequently, we purified the exosomes from the supernatant using a commercial kit. Scanning electron microscopy, transmission electron microscopy, dynamic light scattering, Western blot, and bicinchoninic acid protein assay were used to characterize exosomes. Following that, miR-211 mimic was loaded into exosomes (termed TEXomiR) via a modified calcium chloride procedure. The assessment of miR-211a loading efficiency into exosomes was conducted by analyzing its relative expression. MTT, annexin V/PI, and quantitative real-time polymerase chain reaction were used to measure the proliferation, apoptosis and relative expression of miR-211 target genes, respectively.
Methods
After exposing B16F10 cultured cells to serum-free media for 24 hours, we collected the supernatant. Subsequently, we purified the exosomes from the supernatant using a commercial kit. Scanning electron microscopy, transmission electron microscopy, dynamic light scattering, Western blot, and bicinchoninic acid protein assay were used to characterize exosomes. Following that, miR-211 mimic was loaded into exosomes (termed TEXomiR) via a modified calcium chloride procedure. The assessment of miR-211a loading efficiency into exosomes was conducted by analyzing its relative expression. MTT, annexin V/PI, and quantitative real-time polymerase chain reaction were used to measure the proliferation, apoptosis and relative expression of miR-211 target genes, respectively.
Results
Our study showed that the exosomes can deliver miR-211 mimic efficiently. The treatment of B16F10 cells with miR-211-enriched TEX downregulated miR-211 target genes, including brain-specific homeobox, vascular endothelial growth factor, and transforming growth factor-β receptor. The results indicated the antiproliferative effect of TEXomiR as time-dose-dependent. The flow cytometry evaluation showed that TEXomiR could induce the apoptosis of B16F10 cells. Conclusions: Our data indicated that exosomes can be suitable carriers for miR-211 mimic. Moreover, TEXomiR via anti-cancer effects could inhibit the progression of melanoma cancer.