Inhibition of Wnt5a-induced CCL2 expression and macrophage chemotaxis by SFRP5.

(A) and (B) Real-time PCR and ELISA showed that conditioned medium from SFRP5-positive GES-1 inhibited Wnt5a-induced CCL2 expression in macrophages; the inhibitory effect of the condition medium was abolished when SFRP5 expression was knocked down in GES-1. *P<0.01. (C) and (D) Real-time PCR and ELISA showed that conditioned medium from SFRP5-transfected BGC-803 suppressed Wnt5a-induced CCL2 expression in macrophages. *P<0.01. (E) and (F) Real-time PCR and ELISA showed that rSFRP5 inhibited CCL2 induction by Wnt5a transfection in a dose-dependent manner. *P<0.01. (G) Transwell assay showed that cell migration induced by conditioned medium from Wnt5a-transfected macrophages was suppressed by co-culture of Wnt5a-transfected macrophages with GES-1 or SFRP5-transfected BGC-803. *P<0.01. (H) Transwell assay showed that cell migration induced by conditioned medium from Wnt5a-transfected macrophages was inhibited by pretreatment of macrophages with rSFRP5. *P<0.01. Cont, control; ContV, control vector; WntV, Wnt5a vector; GES, GES-1; SFsiR, SFRP5 siRNA; ContsiR, control siRNA; BGC, BGC-803; SFV, SFRP5 vector; ContV, control vector. Data are expressed as mean±SD, n = 3.