Toad venom induces apoptosis of human osteosarcoma MG-63 cells via down-regulating Integrin α4
Objective It is to explore the effects and possible mechanisms of action of toad venom on proliferation,ap-optosis,invasion and metastasis of human osteosarcoma MG-63 cells.Methods Thirty male C57BL/6J mice were taken to prepare blank serum and different concentrations(2.5%and 5%)of medicated serum containing toad venom.The pas-saged cultured MG-63 cells were divided into 3 groups:the 2.5%toad venom medicated serum group and 5%toad venom medicated serum group were cultured with the corresponding concentrations of toad venom serum,and the control group was cultured with an equal volume of PBS.The proliferative activity of MG-63 cells was detected by CCK-8 assay,the activity of alkaline phosphatase(ALP)in the cells was detected by PNPP assay,the osteogenic ability of cells was detected by aliz-arin red assay,the invasion and metastasis of MG-63 cells were detected by Transwell assay,and the apoptosis rate and the expression of integrin α4 in MG-63 cells were detected by flow cytometry.Results The cell proliferation rates of toad venom medicated serum groups of different concentrations were significantly lower than those of the control group at all time points(all P＜0.05),and the 5%toad venom medicated serum group was significantly lower than that of the 2.5%toad venom medicated serum group at the same period(all P＜0.05);the apoptosis rates and ALP activities of the toad venom medica-ted serum groups of different concentrations were significantly higher than that of the control group(all P＜0.05),and the 5%toad venom medicated serum group was significantly higher than that of the 2.5%toad venom medicated serum group(P＜0.05);Transwell staining results showed that there were orange-red nodules and increased cells with clear boundaries in toad venom medicated serum groups of different concentration;the numbers of invasive cells and metastatic cells in the toad venom medicated serum groups of different concentrations were significantly less than those in the control group(all P＜0.05),and the proportion of Integrin α 4 expression was significantly lower than that in the control group(P＜0.05),and the numbers of invasive cells and metastatic cells in the 5%toad venom medicated serum group were significantly less than those in the 2.5%toad venom medicated serum group(all P＜0.05),and the proportion of Integrin α 4 expression in the 5%toad venom medicated serum group was significantly lower than that in the 2.5%toad venom medicated serum group(P＜0.05).Conclusion Toad venom can promote apoptosis,inhibit proliferation,invasion and metastasis,and improve differentiation and osteogenesis of MG-63 cells,and the mechanism may be related to down-regulating expression of Integrin α4.
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