Effects Of Zoledronic Acid On Osteosarcoma Cell Biological Behaviors

Part 1. Effects of Zoledronic acid on the adhesion, migration and invasion of osteosarcoma cellsObjective: To study the effects of Zoledronic acid on adhesion, migration and invasion of osteosarcoma LM8 cell line.Methods: After LM8 cells were treated with Zoledronic acid, Cell Counting Kit-8 assay was used to assess LM8 cells adhesion and the effect of Zoledronic acid on viability of LM8 cells; their migration and invasion to ECM were measured by wound assay and transwell chamber method respectively.Results: At the concentration of 6.25μmol/L,12.5μmol/L,25μmol/L, Zoledronic acid significantly inhibited LM8 cells adhesion,migration and invasion to the ECM. In addition, the effect of Zoledronic acid on viability of LM8 cells was negligible.Conclusion: Zoledronic acid has anti-adhesion, anti-migration and anti-invasion effects on osteosarcoma LM8 cells in vitro.Part 2. Effects of Zoledronic acid on expression of VEGF in osteosarcoma cellsObjective: To study the effects of Zoledronic acid on expression of VEGF in osteosarcoma LM8 cell line.Methods: After LM8 cells were treated with Zoledronic acid, Semi-quantitative PCR was used to assess the expression of VEGF mRNA; the expression of VEGF protein was detected by immuniohistochemical staining.Results: The transcription of VEGF mRNA and expression of VEGF protein were found in LM8 cells. The transcription of VEGF mRNA and expression of VEGF protein could be down-regulated by treatment with 25μmol/L, 50μmol/L Zoledronic acid for 24h and the effects had a dose dependent relationship(p < 0.05) .Conclusion: Zoledronic acid can inhibit VEGF mRNA transcription and protein expression in osteosarcoma LM8 cell line.Part 3. Effect of zoledronic acid on vasculogenic mimicry in osteosarcoma cells in vitroObjective: To investigate the effect of Zoledronic acid on vasculogenic mimicry formation in osteosarcoma cell line LM8 in vitro.Methods: Osteosarcoma cell line LM8 were tested for their ability to form vasculogenic mimicry formation and the effect of Zoledronic acid on vasculogenic mimicry formation in three-dimensional culture containing type I collagen. The ultrostructure of LM8 cells were determined by electron microscope.Results: Zoledronic acid can inhibit significantly vasculogenic mimicry formation of LM8 cells in vitro, and decrease the surface microvillus of LM8 cells.Conclusion: The mechanism of Zoledronic acid to inhibit vasculogenic mimicry formation of osteosarcoma cells line LM8 is associated with ultrostructure of LM8 cells.