摘要
目的 探讨亚砷酸钠调控wnt信号通路对口腔鳞状细胞癌增殖及凋亡的影响.方法 分别采用125、2.5、5、10、20μmol/L 的亚砷酸钠处理人口腔鳞状细胞癌细胞株Tca8113 24、48、72 h,CCK8 实验检测细胞增殖情况.采用0.10 μmol/L 的亚砷酸钠处理Tca8113 细胞48 h 后,流式细胞仪检测细胞凋亡,Western blot 检测Cleaved caspase3、β-catenin、Cyclin D1 蛋白表达.将Tca8113 细胞分为对照组、亚砷酸钠组(10 μmol/L)、激活剂(10 mmol/L 的LiCl)+亚砷酸钠组(10 μmol/L 亚砷酸钠),各组细胞处理48 h 后,CCK8 实验、流式细胞术及Western blot 分别检测3 组细胞增殖、凋亡及Cleaved caspase3、β-catenin、Cyclin D1 蛋白表达.结果 随着作用时间及亚砷酸钠浓度增加,Tca8113 细胞增殖抑制率均显著升高,具有时间和浓度依赖性(P＜0.05或P＜0.01),根据IC50选择10μmol/L的亚砷酸钠作为后续研究.10μmol/L的亚砷酸钠组细胞凋亡率及Cleaved caspase3蛋白表达显著高于0μmol/L组,β-catenin、Cyclin D1蛋白表达显著低于0μmol/L组(P＜.0.01).亚砷酸钠组及激活剂+亚砷酸钠组细胞凋亡率、细胞抑制率及Cleaved caspase3蛋白表达均显著高于对照组,β-catenin、Cyclin D1蛋白表达显著低于对照组(P＜0.01);激活剂+亚砷酸钠组细胞凋亡率、细胞抑制率及Cleaved caspase3蛋白表达均显著低于亚砷酸钠组,β-catenin、Cyclin D1蛋白表达显著高于亚砷酸钠组(P＜.0.01).结论 亚砷酸钠可抑制口腔鳞状细胞癌增殖及促进凋亡,其机制与Wnt信号通路的调控有关.
Abstract
Objective To investigate the effect of sodium arsenite by Wnt signaling pathway on proliferation and apoptosis of oral squamous cell carcinoma.Methods Cell proliferation was detected after 1.25,2.5,5,10,20μmol/L sodium arsenite treatment human oral squamous cell carcinoma cell line Tca8113 for 24,48,72 hours by CCK8 experiment.0 and 14μmol/L sodium arsenite was used to treatment Tca8113 cells with 48h,cell apoptosis were detected by flow cytometry,Cleaved Caspase3,β-catenin,Cyclin D1 protein expression were detected by Western blot.Tca8113 cells were divided into control group,sodium arsenite group,activating agent+sodium arsenite group,all treated for 48hour,cell proliferation,apoptosis and Cleaved Caspase3,β-catenin,Cyclin D1 protein expression were detected by CCK8 assay,flow cytometry and Western blot.Results Tca8113 cell proliferation was inhibited significantly with the increase of treatment time and sodium arsenite concentration,and has a time and concentration dependent manner(P<0.05 or P<0.01).10μmol/L sodium arsenite as a follow-up study according to the IC50.Cell inhibition rate,apoptosis rate and Cleaved Caspase3 protein expression in 10μmol/L group were significantly higher than that of 0 mol/L group,the expression of β-catenin,Cyclin D1 protein was significantly lower than that of 0 mol/L group(P<0.01).Apoptosis rate,cell inhibition rate and Cleaved Caspase 3 protein expression in sodium arsenite group and activating agent+sodium arsenite group were significantly higher than control group,the expression of β-catenin and Cyclin D1 protein were significantly lower than control group(P<0.01).Apoptosis rate,cell inhibition rate and Cleaved Caspase 3 protein expression in activating agent + sodium arsenite group were significantly lower than that of sodium arsenite group,the expression of β-catenin and Cyclin D1 protein were significantly higher than that of sodium arsenite group(P<0.01).Conclusion Sodium arsenite can inhibit the proliferation of oral squamous cell carcinoma and promote apoptosis,and the mechanism was related to regulation of Wnt signaling pathway.
NETL
NSTL
维普
万方数据