Figure 1

Expression of activated Hh-GLI pathway molecules. Total RNA extracted from osteosarcoma cell lines were used for real-time PCR. Real-time PCR revealed that 4 of 5 human osteosarcoma cell lines increased Sonic Hedgehog (SHH) 2.1- to 18.8-fold (Fig. 1). In addition, 5 of 5 osteosarcoma cell lines increased Desert Hedgehog 1.3- to 24.4-fold (Fig. 1). To further examine Hh pathway molecules expression, we performed real-time PCR for Hh receptors and Hh target genes. PTCH1 was up-regulated 2.7-to 65.8-fold in 5 of 5 human osteosarcoma cell lines. SMO was up-regulated 2.1-to 5.8-fold in 4 of 5 human osteosarcoma cell lines. SMO was up-regulated 2.1-to 5.8-fold in 4 of 5 human osteosarcoma cell lines. GLI1 was up-regulated 2.5-to 8.9-fold in 5 of 5 human osteosarcoma cell lines. GLI2 was up-regulated 1.2-to 9.9-fold in 5 of 5 human osteosarcoma cell lines. The comparative Ct (ΔΔCt) method was used to determine fold change in expression using βII-microglobulin, GAPDH or ACTB. Each sample was run minimally at three concentrations in triplicate (error bar means S.D.). The experiment was triplicate with similar results.