灵芝外泌体样纳米囊泡对乳腺癌细胞增殖、侵袭及迁移的影响OA

Objective To investigate the effect of Ganoderma lucidum exosome-like nanovesicles(GLEVs)on proliferation,invasion,and migration of breast cancer(BC)MCF-7 cells,and to clarify the underlying mechanism by which GLEVs inhibit MCF-7 cell functions.Methods GLEVs were isolated and purified from the supernatant of fresh Ganoderma lucidum using a commercial kit,and the morphology of GLEVs was observed by transmission electron microscopy.MCF-7 cells were treated with 40,80,and 120 μg/mL GLEVs for 24 and 48 h,respectively,and the uptake of GLEVs by MCF-7 cells was detected using the DiO dye phagocytosis assay.MCF-7 cells were divided into blank control group,paclitaxel-positive control group,and GLEVs groups at different concentrations(40,80,120 μg/mL).After 24 and 48 h of culture,the cell proliferation capacity of each group was measured by the MTT assay,and the invasion capacity of MCF-7 cells was detected by the Transwell chamber assay.The migra-tion capacity of MCF-7 cells was evaluated by the Transwell chamber assay after 24 h of culture.Results Under transmission electron microscopy,GLEVs exhibited a typical cup-shaped morphology,indicating successful extraction of GLEVs.DiO staining showed that after 24 h of culture,the uptake of 40,80,and 120 μg/mL GLEVs by MCF-7 cells was higher than that in blank control group,and the uptake gradually increased with rising concentrations.After 48 h of culture,GLEVs at various concentrations were effectively internalized by MCF-7 cells.Notably,at a mass concentration of 120 μg/mL,green fluorescence was uniformly distributed around the cell membrane,exhibiting a concentration-dependent manner.MTT assay results demonstrated that compared with blank control group,after 24 h of culture,the cell proliferation in positive control group was significantly reduced(P<0.001),and that in 40 and 120 μg/mL GLEVs groups was also reduced(P<0.05).Compared with blank control group,after 48 h of culture,the cell proliferation in positive control group was significantly reduced(P<0.000 1),that in 40 and 120 μg/mL GLEVs groups was reduced(P<0.01),and that in 80 μg/mL GLEVs group was significantly reduced(P<0.001).Furthermore,the inhibitory effect on cell proliferation gradually weakened with increasing GLEVs concentration.Cell invasion capacity was evaluated by Transwell assay.Com-pared with blank control group,after 24 h of culture,the invasion capacity of 40,80 and 120 μg/mL GLEVs groups was significantly reduced(P<0.01,P<0.05).Transwell assay results indicated that compared with blank control group,80 μg/mL GLEVs group could significantly inhibit the migration capacity of MCF-7 cells(P<0.05).Conclusion GLEVs can be internalized by MCF-7 cells and inhibit the proliferation,invasion,and migration of MCF-7 cells.