Fig. 4
Exosomal transfer of miR-99a-5p to human peritoneal mesothelial cells (HPMCs) promotes EOC cell invasion. a miRNA qRT-PCR. Relative expression levels of miR-99a-5p in HPMCs treated with IOSE- and EOC-derived exosomes at 100 μg/mL for 24 h are shown. Expression levels of miR-99a-5p in HPMCs treated with PBS alone were set to 1.0. Data represent the mean ± SD of three experiments. b A schematic of the HPMC-coated in vitro invasion assay. HPMCs were plated onto 24-well cell culture inserts coated with Matrigel. After HPMCs reached confluence, cancer cells were incubated on the HPMCs monolayer and allowed to invade for 24 h. Cells that migrated to the trans-side of the membrane were quantitated. c HPMCs were pre-cultured with TYK-nu-derived exosomes at 100 μg/mL for 24 h and then TYK-nu cells were plated onto the HPMC monolayer. Twenty-four hours after incubation, invading cells on the underside of the filter were counted. Representative pictures of the chambers are shown below. d miRNA qRT-PCR. Relative expression levels of miR-99a-5p in HPMCs transfected with miR-99a-5p are shown. e In vitro cell proliferation assay. HPMCs were transfected with negative control miRNA or precursor miR-99a-5p. Twenty-four hours later, relative cell proliferation was assessed using the MTS assay. f HPMC-coated in vitro invasion assay. After HPMCs were transfected with negative control miRNA or precursor miR-99a-5p, TYK-nu cells were plated onto the HPMC monolayer and allowed to invade. g miRNA qRT-PCR. Relative expression levels of miR-99a-5p in HPMCs transfected with anti-miR-99a-5p are shown. h In vitro cell proliferation assay. HPMCs were transfected with negative control miRNA and anti-miR-99a-5p. i HPMC-coated in vitro invasion assay. After HPMCs transfected with negative control miRNA or precursor anti-miR-99a-5p were pre-cultured with TYK-exo, TYK-nu cells were plated onto the HPMC monolayer and allowed to invade. Data represent the mean ± SD of three experiments. *p < 0.05, **p < 0.01, and ***p < 0.001. n.s., not significant