国际肿瘤学杂志

国际肿瘤学杂志 ›› 2017, Vol. 44 ›› Issue (2): 86-90.doi: 10.3760/cma.j.issn.1673-422X.2017.02.002

• 论著 • 上一篇    下一篇

吴茱萸碱抑制Wnt/β-catenin信号通路诱导骨肉瘤MG-63细胞凋亡

袁源,李松霖,王忠华,沈会华,李悟,王卫东   

  1. 830000 乌鲁木齐，解放军新疆军区总医院心胸外科（袁源、王忠华、沈会华、李悟、王卫东）；第三军医大学附属大坪医院野战外科研究所肿瘤中心（李松霖）
  • 出版日期:2017-02-08 发布日期:2017-01-04
  • 通讯作者: 王忠华，Email: yyccr@hotmail.com E-mail:yyccr@hotmail.com

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

Yuan Yuan, Li Songlin, Wang Zhonghua, Shen Huihua, Li Wu, Wang Weidong   

  1. Department of Cardio Thoracic Surgery, Xinjiang Military Region General Hospital, PLA, Urumqi 830000, China
  • Online:2017-02-08 Published:2017-01-04
  • Contact: Wang Zhonghua E-mail:yyccr@hotmail.com

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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24 h，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、0.5 μmol/L、1.0 μmol/L、2.0 μmol/L和4.0 μmol/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。

关键词: 吴茱萸碱, 骨肉瘤, 细胞增殖, 细胞凋亡, Wnt/β-catenin

Abstract: Objective To investigate the effects and mechanism of evodiamine on the proliferation and apoptosis of osteosarcoma MG63 cells. MethodsMG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. ResultsAs 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