吴茱萸碱抑制Wnt/β-catenin信号通路诱导骨肉瘤MG-63细胞凋亡

Objective To investigate the effects and mechanism of evodiamine on the proliferation and apoptosis of osteosarcoma MG-63 cells. Methods MG-63 cells were cultured with evodiamine for 24 hours, and the cell proliferation was evaluated by methyl thiazolyl tetrazolium (MTT) assay. Cell cycle arrest, apoptosis and intracellular Ca2+ accumulation were evaluated by flow cytometry. BALB/C mice model of osteosarcoma was established to investigate the tumor inhibitory effect of evodiamine on human osteosarcoma. Wnt/β-catenin signaling protein expressions in osteosarcoma cells were detected by Western blotting. Results As concentration of evodiamine increasing (0.25 μmol/L, 0.5 μmol/L, 1.0 μmol/L, 2.0 μmol/L and 4.0 μmol/L), the inhibition rate of MG-63 cells increased [(4.18±1.26)%, (15.49±2.26)%, (40.55±6.57)%, (49.87±7.69)% and (60.42±8.29)%]. The difference was statistically significant between 2.0 μmol/L group and the control group (t=-2.66, P<0.05). MG-63 cells were cultured with 2.0 μmol/L evodiamine for 24 hours, and the apoptotic rate was (64.67±8.63)%, the proportion of S phase cells was (85.33±9.31)%, the fluorescence of Ca2+ was 97.33±21.31. The corresponding data of the control group were (4.94±0.81)%, (43.67±8.92)% and 28.67±8.92, the differences were statistically significant (t=-11.90, P<0.05; t=-7.22, P<0.05; t=-6.65, P<0.05). On mice model, the tumor weight of evodiamine group and the control group was (2.15±0.35)g and (4.29±0.49)g respectively, the difference was statistically significant (t=7.95, P<0.05). Comparing with the control group (1.00±0.00), evodiamine decreased the expression of β-catenin protein (0.72±0.36) and increased the expressions of Bax (1.15±0.27) and Caspase-3 (1.46±0.18) protein, and the differences were statistically significant (t=-3.05, P<0.05; t=-6.42, P<0.05; t=-5.85, P<0.05). Conclusion Evodiamine inhibits proliferation and induces apoptosis of human osteosarcoma MG-63 cells by blocking Wnt/β-catenin signaling. Key words: Evodiamine; Osteosarcoma; Cell proliferation; Apoptosis; Wnt/β-catenin