Gr127935

The drugs consisted of lisuride maleate (#4052), clozapine (#0444), SCH23390 (#0925), GR127935 (#1477; Bio-Techne Corporation, Minneapolis, MN), LSD-tartrate (NIDA Drug Supply Program, Bethesda, MD), fluoxetine HCl (#F132) and reserpine (#R0875; Sigma-Aldrich, St. Louis, MO). The vehicle for lisuride, LSD, MDL, reserpine, SCH23390, and GR127935 consisted of N,N-dimethyllacetamide (final volume 0.5%; Sigma-Aldrich) and it was brought to volume with 5% 2-hydroxypropoyl-β-cyclodextrin (Sigma-Aldrich) in sterile water (Mediatech Inc., Manassas, VA). clozapine was solubilized with glacial acetic acid (0.05% final) and was brought to volume with 5% 2-hydroxypropyl-β-cyclodextrin in sterile water. Fluoxetine was solubilized in sterile water. All drugs were injected (i.p.) in a 5 ml/kg volume.

The human colorectal cancer LoVo, HCT-116, HT-29 and SW403 cell lines were purchased from the Shanghai Cell Collection (Shanghai, China). Cells were grown in RPMI 1640 medium supplemented with 10% (v/v) heat-inactivated fetal calf serum, 2 mM glutamine, 100 units/ml penicillin, and 100 μg/ml streptomycin (Invitrogen, Carlsbad, CA) at 37°C in a 5% CO₂-humidified atmosphere. Monoclonal antibodies against 5-HT family, Axin1, β-catenin, APC, GSK3β, CKIα, LEF1, TCF4, cyclin D1, c-Myc, MMP-2, MMP-9, MMP-7 and β-actin were products of Cell Signaling Technology (Beverly, MA, USA). 5-HT_1DR agonist (Sumatriptan) and 5-HT_1DR antagonist (GR127935) were from Sigma Chemical Co. (St. Louis, MO).