Tetrandrine suppresses the growth of human osteosarcoma cells by regulating multiple signaling pathways

Abstract

Although osteosarcoma (OS) is the most common malignant tumor among juvenile bone tumors, its treatment plan and clinical outcome have not improved significantly in recent decades. Tetrandrine (TET), a Chinese medicine that is usually used in the therapy of silicosis, hypertension and arthritis, has been confirmed by many studies to possess potent antitumour growth properties, but there are different limitations when describing specific mechanisms. Here, we found that TET can obviously prevent the proliferation, migration and invasion of both 143B and MG63 cells and promote their apoptosis in vitro. Our results for luciferase reporter and Western blotting assays show that TET may exert its antitumour activity by regulating multiplex signaling pathways, including the MAPK/Erk, PTEN/Akt, Juk and Wnt signaling pathways. Furthermore, the regulatory effect of TET on OS cells and related signaling pathways was verified again in vivo. Our findings suggest that the anticancer function of TET on human OS may be mediated by its targeting of multiple signaling pathways and that TET may be used as a single drug or in combination with other drugs during the treatment of OS.

Epigallocatechin-3-gallate suppresses the growth of human osteosarcoma by inhibiting the Wnt/β-catenin signalling pathway

“…After treatment with EGCG for 24 hours, the cells were gently washed three times with PBS and then fixed with 4% paraformaldehyde for 10 minutes at 37°C. After the paraformaldehyde was discarded, the cells were gently washed three times with PBS and then stained with Hoechst 33,258 dye for 5 minutes in the dark [ 18 ]. The nuclear morphology was analyzed by fluorescence microscopy (Eclipse Ti, Nikon, Japan).…”

“…A microplate reader was then used to detect the absorbance value of the culture medium at 492 nm. The OD values at different time points were converted using the blank group value as the 100% reference point, and the converted results were used for statistical analysis of cell proliferative activity [ 18 ].…”

“…First, 143B cells were inoculated in 6-well plates, and after the cell confluence reached 60%, the cells were treated with EGCG for 24 hours. The cells were collected at the designated time, washed repeatedly with PBS, and then resuspended in 500 µl of PBS [ 18 ]. The distribution of cells in early and late apoptosis was detected by flow cytometry using an Annexin V-FITC/PI double labeling staining kit (Beyotime Biotechnology, Shanghai, China).…”

“…The upper Transwell chamber (Corning Corporation, USA) was seeded with approximately 20,000 cells per well and 200 µl of culture medium with 10% FBS, and 600 µl of culture medium (10% FBS) containing different concentrations of EGCG was added to the lower Transwell chamber. After culture for 24 hours, the cells that migrated to the lower chamber were fixed with 4% paraformaldehyde and then stained with 0.1% crystal violet [ 18 ].…”

“…The Transwells were placed in an oven at a constant temperature of 37°C for approximately 2 hours until gels formed. The next experimental steps were the same as those in the Transwell migration assay [ 18 ].…”