Journal: Scientific Reports
Article Title: Polyphyllin I suppresses human osteosarcoma growth by inactivation of Wnt/β-catenin pathway in vitro and in vivo
Figure Lengend Snippet: PPI suppressed osteosarcoma cells by specifically inactivating Wnt/β-catenin signaling pathway. ( A ) RT-PCR analysis of β-catenin expression level in matched human osteosarcoma tissues (tumors) and adjacent noncancerous tissues (normal) from 3 patients. ( B ) 143-B cells and ( C ) HOS cells were respectively treated with 0.8 μM PPI for indicated times, and expressions of test proteins were examined by western blotting analysis, β-actin was used as loading control, and the full-length blots were included in the supplementary information file as Figures and . 143-B cells were pretreated with 4 μM CHIR9902 (the specific GSK-3β inhibitor) for 24 h before exposed to 0.8 μM PPI for another 48 h, the combined CHIR and PPI treatments result in rescued ( D ) active β-catenin expression (the full-length blots were included in the supplementary information file as Figure ) and cell viability ( E ) compared to the PPI treatment alone in 143-B osteosarcoma cells. 143-B cells were transfected with either small interfering RNA-targeting β-catenin (si-β-catenin) or si-control for 48 h before exposed to 0.8 μM PPI for another 24 h, si-β-catenin potentiated PPI induced ( F ) down-regulation of active β-catenin expression (the full-length blots were included in the supplementary information file as Figure ), ( G ) decrease of cell viability and ( H ) inhibition of migration of 143-B osteosarcoma cells induced by PPI. * p < 0.05, ** p < 0.01, *** p < 0.001, versus the control.
Article Snippet: Briefly, 143-B cells were co-transfected with either small interfering RNA-targeting β-catenin (100 nM si-β-catenin) or 100 nM si-control for 72 h, using Lipofectamine 2000 (Invitrogen). β-catenin siRNA II (#6238) was bought from Cell Signaling Technology (Danvers, USA). si-control was designed and produced by Shanghai GenePharma Co.,Ltd (Shanghai, China).
Techniques: Reverse Transcription Polymerase Chain Reaction, Expressing, Western Blot, Transfection, Small Interfering RNA, Inhibition, Migration