Cell invasion assays
HuCCA-1 or RMCCA-1 cells (5x104 cells) were
plated in Matrigel®-coated transwell (Costar, 8 μm pore
sized) plates and incubated overnight at 37˚C in 5% CO2
incubator. The cells were treated with 10 and 20μg/ml
(ethanol extract) or with 2.5 and 5μg/ml (KP.8.10
compound), respectively, in 200 μl of serum-free medium
containing 0.1% BSA for 30 min. The same
concentrations of the extracts, with or without 20 ng/ml
of rhHGF, were added into the lower chamber. The number
of invaded cells was counted after 24 or 48 h incubation,
under the microscope (40x magnification), for 6 random
fields per membrane and averaged.