The Experimental Study Of The Effect Of ⁹⁹Tc-MDP On Osteosarcoma MG63 Cells

Objective:The aim of this study was to investigate the effect and its related mechanisms of technetium methylene diphosphonate("Tc-MDP)on Osteosarcoma(OS)MG63 cells.It will provide a theoretical basis for clinical treatment of OS.Methods:Different concentrations of 99Tc-MDP were applied to MG63 cells.Cell proliferation inhibition rate was detected by the method of Cell counting kit-8(CCK-8).Cell apoptosis and cell cycle were analyzed by flow cytometry.Immunocytochemistry(ICC)and western blot(WB)were used to evaluate the proteins expression level of Osteoprotegerin(OPG),Receptor activator of NF-κB(RANK)and Receptor activator of NF-κB Ligand(RANKL).Results:1.Compared with the control group,100μg/mL 99Tc-MDP slightly promoted the proliferation of MG63 cells.But with the extension of time,the drug action turns into inhibition.The difference was statistically significant(P<0.01).The CCK-8 method showed that in the range of(200,300,400,500,600)μg/mL of the 99Tc-MDP had a concentration-dependent dose-dependent effect on the MG63 cells(P<0.01).2.The apoptotic rates of the control group,100μg/mL group and 400pg/mL group were 0.187±0.061%,10.507±1.092%and 22.787±0.556%,respectively.99Tc-MDP can promote the apoptosis of MG63 cells.And the differences were statistically significant(P<0.01).3.The effect of 99Tc-MDP on its cycle mainly showed G1 arrest.And there was dose-dependent.There was no significant difference between the control group and the 100μg/mL 99Tc-MDP concentration group(P>0.05).The difference between 100μg/mL and 400μg/mL 99Tc-MDP concentration group was statistically significant(P<0.01).4.The related protein OPG was found in MG63 cells,which was mainly expressed in cytoplasm.Compared with the control group,100μg/mL 99Tc-MDP and 400μg/mL 99Tc-MDP decreased the expression of OPG in MG63 cells.And the differences were statistically significant(P<0.01).5.Compared with the control group,the expression of OPG in MG63 cells was inhibited by 100μg/mL 99Tc-MDP and 400μg/mL99Tc-MDP.The former difference was not statistically significant(P>0.05).But latter difference was statistically significant(P<0.01).Compared with the control group,100μg/mL 99Tc-MDP had a promoting effect on the expression of RANK in MG63 cells.400μg/mL99Tc-MDP had an inhibitory effect on the expression of RANK in MG63 cells.And the differences were statistically significant(P<0.01).Compared with the control group,100μg/ml 99Tc-MDP inhibited the expression of RANKL in MG63 cells and the difference was statistically significant(P<0.01).The expression of RANKL in MG63 cells by 400μg/ml 99Tc-MDP was a promoting effect.And the difference was not statistically significant(P>0.05).Compared with the control group,100μg/mL 99Tc-MDP decreased the expression ratio of OPG/RANK in MG63 cells.And the ratio of OPG/RANK expression in MG63 cells increased by 400μg/mL 99Tc-MDP.The difference was statistically significant.(P<0.01).Conclusions:High-concentration 99Tc-MDP can inhibit the proliferation of MG63 cells,while the low-concentration 99Tc-MDP can slightly promote the proliferation of MG63 cells in the early stage.But the effect was prolonged with the prolonged action time.It has concentration and time dependence.High-concentration of 99Tc-MDP induced apoptosis and cycle arrest in MG63 cells.The related protein OPG is present in MG63 cells,which is mainly expressed in the cytoplasm.High-concentration of 99Tc-MDP can effectively inhibit the expression of RANK in MG63 cells.