Figure 3

Inhibition of Hh pathway prevents osteosarcoma growth in vitro. A, We performed real-time PCR to determine which concentration of cyclopamine effectively inhibited Hh-GLI activity in osteosarcoma cells, and then measured the expression of the Hh-GLI pathway target PTCH1 and GLI2. Cyclopamine at 20 μM reduced mRNA levels of PTCH1 in 143B cell (error bar means S.D.). The comparative Ct (ΔΔCt) method was used to determine fold change in expression using ACTB. Each sample was run minimally at three concentrations in triplicate (error bar means S.D.). The experiment was triplicate with similar results. B, Growth of viable 143B and HOS cells over 3 days was slowed in dose-dependent fashion by cyclopamine treatment. The experiment was triplicate with similar results. C, Colony formation assay revealed cyclopamine reduced colony formation in soft agar. The experiment was triplicate with similar results. (*: P < 0.01) (error bar means S.D.)