Figure 4.

Ovatodiolide treatment resulted in less oncogenic exosomes produced by colon cancer tumorspheres. Exosomes, isolated from tumorspheres generated from ovatodiolide-treated (Exo^sp_OV) HCT116 and HT29, showed a significantly reduced ability to promote tumorsphere formation (A), 5-FU resistance (B), and migration (C) when co-cultured with parental HCT116 and HT29 cells. (D) Exo^sp_OV cultured normal fibroblasts (NF) showed a lower a-SMA expression (upper panel) and reduced amount of IL-6 released (lower panel) as compared to their Exo^sp-cultured counterparts. (E) Real-time PCR analysis of M1 M2 markers post macrophages co-cultured with Exo^sp and Exo^sp_OV (upper panel). Comparative ELISA of TGF-β1 released by macrophages after co-cultured with Exo^sp and Exo^sp_OV (lower panel). MΦ, uncommitted macrophages; M1 TAM markers, TNF-α, IL-1β; M2 TAM markers, CD206, TGF-β1. a, P<0.001; b, P<0.01; NS, no significant difference (all compared to MΦ). *P<0.05; **P<0.01; ***P<0.001.