Effect And Mechanism Of Cistanche On Reversing Chemotherapy-resistant Of Osteosarcoma(MG-63) Cells

Objective: In vitro experiments were conducted to study the reversal effect of Cistanche desertiola on MG-63/MTX resistant cells in human osteosarcoma,and to explore the mechanism by which cistanche reversed MG-63/MTX cells.Methods: Three groups of Cistanche deserticolata drug-containing serum and control g roup serum were obtained by gavage of SD rats.Detect the IC50 value of MTX befor e drug intervention with MTT method,MG-63 cells were divided into four groups,respe ctively,high dose group(A),middle dose group(B),low dose group(C),saline group(N),Usi ng IC50 value of MTX intervention each grop cells,replace the medium after 24 hours and adding different concentrations of drug-containing serum containing medicine seru m and blank serum,The intervention and medium replacing was continued until the co ntrol group cells could stable growth with IC50 concentration of MTX.Observe each c ell morphological changes,determined by MTT method is used to detect and and calcu late the IC50 and resistance index after drug intervention,detect the expression of MR P1 and the rate of apoptosis with flow cytometry,detect the expression of P53 protein in each group with Western blot.Results:(1)MG-63 cells grow adherently,with medium volume and uniform size.The c ells are spindle-shaped or irregular,with pseudopods,less cytoplasm,larger nucleus,an d clear nucleoli.After drug intervention,the cells in groups A,B,C,and N increased in volume compared with the original MG-63;(2)The IC50 values of the A,B,C and N groups of cells after MTX intervention were:(4.60±0.14)ug/ml、(6.32±0.30)ug/ml、(7.04±0.14)ug/ml、(9.28±0.18)ug/ml,Resistance index were:1.80、2.55、2.84、3.74;(3)Apo ptosis rates of A,B,C and N groups of cells were(39.30±1.85)%,(29.33±1.65)%,(24.33±0.91)%,(18.23±1.51)%,A>B>C>N,There was a statistical difference between the fou r groups(P<0.01);(4)After the intervention,the expressions of MRP1 protein in the four groups A,B,C and N were:(8.47±0.91)%,(13.80±1.2)%,(28.97±1.2)%,(39.77±1.81)%,A<B<C<N,The four groups are statistically significant(P<0.001);(5)Compared with the gro up N,the expression of P53 protein in each of the groups A,B and C was down-regul ated,Group A<Group B<Group C<Group N,There are statistical differences betweenthe groups(P<0.05).Conclusion:Cistanche deserticolata can reverse the drug-resistant of Osteosarcoma MG-63/MTX.The mechanism may be related with induce apoptosis and inhibit protein expr ession of MRP1 and P53 of MG-63/MTX.