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. 2023 Nov 16;115(1):155–169. doi: 10.1111/cas.16014

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© 2023 The Authors. Cancer Science published by John Wiley & Sons Australia, Ltd on behalf of Japanese Cancer Association.

This is an open access article under the terms of the http://creativecommons.org/licenses/by-nc-nd/4.0/ License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non‐commercial and no modifications or adaptations are made.

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MicroRNA (miR)‐296‐3p is specifically upregulated in activated cancer‐associated fibroblast (CAF)‐derived extracellular vesicles (EVs) from ovarian cancer. (A) Volcano plot showing the differentially expressed miRNAs between quiescent CAF (qCAF)‐ and activated CAF (aCAF)‐derived EVs. (B) Real‐time PCR analysis of miR‐296‐3p expression in qCAFs, aCAFs, and their respective EVs. (C) dbDEMC database analysis of miR‐296‐3p levels in normal ovarian tissues and ovarian cancer tissues. (D) Bioinformatics analysis of miR‐296‐3p expression in ovarian cancer based on tumor stages using the UALCAN database. (E) Real‐time PCR showing the expression of miR‐296‐3p in normal ovarian epithelial cells (HOSE 6‐3 and HOSE 17‐1), ovarian cancer cells (A2780, SKOV3, HO8910, OVCAR5, OV90, CAOV3, and ES‐2), normal fibroblasts (NFs), and CAFs. (F) CAF markers, including α‐smooth muscle actin (α‐SMA), vimentin, and FAP, were determined in three pairs of NFs and CAFs in colorectal cancer by real‐time PCR. (G) Real‐time PCR showing the expression of miR‐296‐3p in three pairs of NFs and CAFs in colorectal cancer. (H) EVmiRNA database analysis of miR‐296‐3p expression in EVs from different sample sources. Red arrows indicate the miR‐296‐3p expression in EVs from fibroblasts. (I) Real‐time PCR showing the expression of miR‐296‐3p in NFs, CAFs, and their respective EVs in ovarian cancer. (J) Relative expression of miR‐296‐3p in tumor cells after incubation with blank or qCAF‐ or aCAF‐derived EVs for 24 or 48 h. *p < 0.05; **p < 0.01; ***p < 0.001; ****p < 0.0001. ns, no significance; RPM, reads per million; TCGA, The Cancer Genome Atlas.