吴茱萸碱抑制Wnt/β-catenin信号通路诱导骨肉瘤MG-63细胞凋亡  

Evodiamine inhibits apoptosis of human osteosarcoma MG-63 cells by blocking Wnt/β-catenin signaling

作　　者：袁源[1] 李松霖[2] 王忠华[1] 沈会华[1] 李悟[1] 王卫东[1] 

机构地区：[1]解放军新疆军区总医院心胸外科,乌鲁木齐830000 [2]第三军医大学附属大坪医院野战外科研究所肿瘤中心

出　　处：《国际肿瘤学杂志》2017年第2期86-90,共5页Journal of International Oncology

摘　　要：目的 探讨吴茱萸碱对骨肉瘤MG-63细胞增殖与凋亡的影响及其机制.方法 吴茱萸碱作用于骨肉瘤MG-63细胞24h,四甲基偶氮唑蓝（MTT）法检测吴茱萸碱对MG-63细胞的增殖抑制作用.流式细胞术检测吴茱萸碱对细胞周期、细胞凋亡及细胞内钙离子浓度的影响.利用BALB/C小鼠建立骨肉瘤移植瘤模型,观察吴茱萸碱对骨肉瘤模型的生长影响.用Western blotting检测吴茱萸碱对Wnt/β-catenin信号通路蛋白表达的影响.结果 随着吴茱萸碱浓度（0.25 μmol/L、0.5μmol/L、1.0 μmol/L、2.0μmol/L和4.0μmol/L）增加,MG-63细胞抑制率也明显增加[（4.18±1.26）％、（15.49 ±2.26）％、（40.55±6.57）％、（49.87 ±7.69）％和（60.42±8.29）％],2.0 μmol/L组与对照组（0）比较差异有统计学意义（t=-2.66,P＜0.05）.2.0 μmol/L吴茱萸碱作用MG-63细胞24 h后凋亡率为（64.67±8.63）％、S期细胞比例为（85.33±9.31）％、钙离子荧光值为97.33±21.31,对照组凋亡率为（4.94±0.81）％、S期细胞比例为（43.67±8.92）％、钙离子荧光值为28.67±8.92,差异均有统计学意义（t=-11.90,P＜0.05;t=-7.22,P＜0.05;t=-6.65,P＜0.05）.小鼠模型吴茱萸碱组肿瘤重量为（2.15 ±0.35）g,对照组肿瘤重量为（4.29±0.49）g,差异有统计学意义（t=7.95,P＜0.05）.与对照组（1.00 ±0.00）比较,吴茱萸碱可降低β-catenin蛋白表达（0.72±0.36）,升高Bax（1.15 ±0.27）、Caspase-3（1.46±0.18）蛋白表达,差异均有统计学意义（t=-3.05,P＜0.05;t=-6.42,P＜0.05;t=-5.85,P＜0.05）.结论 吴茱萸碱可以通过抑制Wnt/β-catenin信号通路抑制骨肉瘤MG-63细胞的增殖并诱导其凋亡.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 ceils 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 ap

关 键 词：吴茱萸碱 骨肉瘤 细胞增殖 细胞凋亡 

分 类 号：R285[医药卫生—中药学]