Global analysis of genes regulated by canonical Wnt pathway

Up-regulation of canonical Wnt signaling has been implicated in liver fibrosis and cancer. To determine canonical Wnt target genes potentially implicated in these diseases, we performed microarray analysis of culture-activated rat hepatic stellate cells (HSCs) treated with canonical Wnt inhibitors, DKK-1, ICG-001, or FJ9. DKK-1, ICG-001, and FJ9 treatment of HSCs: Fully activated primary rat HSCs (cultured for 7 days on plastic dish with 10% FBS containing low glucose DMEM) were treated with: 1) adenovirus expressing DKK-1 (Ad.DKK-1) vs. Ad.GFP (control) at a multiplicity of infection (MOI) of 200 for 24 h in low glucose DMEM with 5% FBS and antibiotics, and then maintained for an additional 24 h in serum-free medium; 2) ICG-001 (10 µM) vs. vehicle for 24 h in serum-free medium; or 3) FJ9 (200 µM) vs. vehicle for 24 h in serum-free medium. No replicates were used. Instead, each of the 6 conditions (3 treatments and 3 respective controls)/samples consisted of a pool of RNA from 3 to 4 rats.