Fig. 5: Identifying unique EV-miRNA profiles of PROC.

a Study design to determine EV-miRNA profiles based on initial surgical tissue samples and ascites of HGSOC. b Characterization of the EVs from patients’ ascites. Nanoparticle tracking analyses demonstrate the particle size of the EVs. Transmission electron microscopy was utilized to visualize the EVs. The scale bar indicates 100 nm. Immunoblot analysis for CD9, CD63, CD81, and GRP of representative EV samples from patient ascites. c Hierarchical clustering and heatmap showing 397 differentially expressed miRNA in ascites-EV between platinum-sensitive and platinum-resistant. The differentially expressed miRNA was defined as an absolute log2 fold change exceeding 1. Volcano plot demonstrating significantly differentially expressed miRNA in ascites-EV between platinum-resistant and platinum-sensitive, based on a cut-off of |log2FC| of >1 and an adjusted P-value of <0.05. Adjusted P-values for each miRNA were calculated using the Wald test in DESeq2. d Principal component analysis of miRNA sequencing in ascites-EV, including 6 platinum-resistant and 6 platinum-sensitive samples. e Hierarchical clustering and heatmap illustrating 385 differentially expressed miRNA in HGSOC tissues between platinum-sensitive and resistant. The differentially expressed miRNA was defined as an absolute log2 fold change of >1. Volcano plot illustrating significantly differentially expressed miRNA in ascites-EV between platinum-resistant and platinum-sensitive, based on a cut-off of |log2FC| of >1 and an adjusted P-value of <0.05. Adjusted P-values for each miRNA were calculated using the Wald test in DESeq2. f Principal component analysis of miRNA sequencing in HGSOC tissues, including six platinum-resistant and six platinum-sensitive samples. g Venn diagram based on volcano plots (c) and (e) demonstrating miRNAs with high expression in both resistant ascites-EV and resistant tissues. h Correlation of miRNA expression in ovarian cancer tissues and ascites-EV based on miRNA sequencing data. R-values were calculated from Spearman’s correlations. i Cisplatin sensitivity of transfected KURAMOCHI and PEO1 cells measured using the MTS assay. KURAMOCHI cells were transfected with a 20-nM miRNA mimic for 24 h, and subsequently, cells were treated with a cisplatin-containing medium for 48 h. PEO1 cells were transfected with a 20 nM miRNA mimic for 24 h, and subsequently, cells were treated with a cisplatin-containing medium for 72 h.