The Study Of Adiponectin Regulating AMPK And ACC Phosphorylation And Cell Proliferation Of Osteosarcoma MG-63Cells

Objective:In this study,we investigate adenpsine monophosphate-activated protein kinase (Adenosine monophosphate activated protein kinase, AMPK), phosphorylation of acetyl coenzyme A carboxylase (-Acetyl carboxylase, ACC),and cell proliferation of the MG-63cells in the status of human adiponectin recombinant.Methods:Select the MG-63cells, SaOS-2cells, hFOB1.19cells,wich is in logarithmic growth phase, then try to get the protein of the cells,and detect the levels of adiponectin receptor1by Western Blot Test. Osteosarcoma cell line MG-63which in logarithmic growth phase were added to5ml mixted adiponectin medium (the concentration is lug/ml), to deal with different time, and detect the levels of AMPK, ACC phosphorylation by Western Blot Test, then choose the best adiponectin in processing time; according to the the adiponectin best processing time, the MG-63cells were exposed in a series concentrations of adiponectin detect the levels of AMPK and ACC phosphorylation by Western Blot Test, determine adiponectin best action concentrations. Based on the optimal duration and optimal concentration and PBS as a control group, the corresponding concentrations of adiponectin for the treatment group, detect the levels of AMPK and ACC phosphorylation by Western blot Test, to study the adiponectin regulating AMPK and ACC phosphorylation of osteosarcoma MG-63cells.Then learn the affection of adiponectin on the MG-63cell proliferation by CCK-8assay.Results:Compared to the normal osteoblasts-hFOB1.19,the levels of AdipoRl in MG-63cells and SaOS-2cells significantly increased (the MG-63cells:P=0.001; SaOS-2cells:P=0.001); the adiponectin on the optimal role of the MG-63cells time is30min (P=0.005), the best role of the concentration is0.1ug/ml (P=0.007), compared with the control group, adiponectin group can significantly promote the AMPK (P=0.001), ACC (P=0.001) phosphorylation of MG-63cells; with a short period of time (24h),the adiponectin on the MG-63cell proliferation was no significantly inhibited (P>0.05).Conclusion:The level of AdipoRl expression in osteosarcoma cells is higher than that in normal osteoblasts.The adiponectin can enhance the phosphorylation of AMPK and ACC; a short time of adiponectin on the proliferation of MG-63cells was no significantly inhibited.