Mechanism Of Total Flavonoids From Epimedii Folium Combined With Firarubicin Upon Osteosarcoma MG-63 Cells

Objective : To investigate the mechanism of total flavonoids from Epimedii Folium(EF) combined with pirarubicin(THP) upon osteosarcoma cells MG-63 in vitro.Methods:Four groups were set: blank control group, EF group(concentration of EF: 50, 100, 200, 400, 800 μg·m L-1), THP group(concentration of THP: 1, 2, 4, 8, 16μg·m L-1) and combination group(concentration of EF+THP: 50+1, 100+2, 200+4,400+8, 800+16 μg·m L-1). Cell morphologies of each groups were observed under inverted phase contrast microscope. Cell inhibition rates of different concentrations were measured by CCK-8. The combination index(CI) of the two drugs was calculated and its interactions were analyzed through Chou-Talalay Method. Cell apoptosis rates of each groups were detected by Annexin V-FITC/PI double staining flow cytometry. Expressions of Caspase-3, P21 and Bcl-2 m RNA were detected by RT-PCR and expressions of caspase-3, p21 and bcl-2 protein were tested by Western blotting.Results:Experiment groups presented apoptosis states in different degrees. Cell proliferation inhibition rates of each group increased along with the increasing of drug concentration. Cell proliferation inhibition and apoptosis rate of combination group were significantly higher than either of the two single drug groups(P<0.05). CI stayed at 0.374~0.770 when combination group inhibition rate from 10% to 90%.Both EF and THP could upregulate the expressions of Caspase-3, P21 m RNA and cleaved-caspase-3, p21 protein. However, Both EF and THP could downregulate the expressions of Bcl-2 m RNA and bcl-2 protein. The trend of combination group was more obvious(P<0.05).Conclution:EF and THP could inhibit osteosarcoma MG-63 cells proliferation significantly, and induced cell apoptosis by upregulating the expressions of Caspase-3,P21 m RNA/protein and downregulating the expressions of Bcl-2 m RNA/protein. It showed synergistic effect when two drugs combined.