基于Wnt/β-catenin通路探讨五味子素B对人舌鳞癌细胞化疗敏感性的作用

Effect of Schisandrin B on Chemotherapeutic Sensitivity of Human Tongue Squamous Cell Carcinoma Based on Wnt/β-catenin Pathway

目的:探讨五味子素B对人舌鳞癌细胞纳武单抗化疗敏感性的作用及对分泌型糖蛋白(Wnt)/β-连环蛋白(β-catenin)信号通路的影响。方法:将对数生长期的人舌鳞癌SCC15细胞随机分为对照组、纳武单抗组、纳武单抗+五味子素B组、纳武单抗+激动剂组、纳武单抗+激动剂+五味子素B组。各组细胞给予相应药物处理后,MTT法检测细胞存活率,流式细胞仪检测细胞凋亡率,划痕实验检测划痕愈合面积百分比,Transwell实验检测侵袭细胞数,蛋白印迹法检测细胞周期蛋白D1(cyclin D1)、原癌基因(c-myc)、β-catenin蛋白相对表达量。结果:与对照组比较,纳武单抗组细胞存活率降低,划痕愈合面积百分比减小,侵袭细胞数减少,cyclin D1、c-myc、β-catenin蛋白相对表达量降低(P<0.05);与纳武单抗组比较,纳武单抗+五味子素B组细胞存活率降低,细胞凋亡率升高,划痕愈合面积百分比减小,侵袭细胞数减少,cyclin D1、c-myc、β-catenin蛋白相对表达量降低(P<0.05),而纳武单抗+激动剂组细胞存活率升高,细胞凋亡率降低,划痕愈合面积百分比增加,侵袭细胞数增多,cyclin D1、c-myc、β-catenin蛋白相对表达量升高(P<0.05);与纳武单抗+五味子素B组比较,纳武单抗+激动剂+五味子素B组细胞存活率升高,细胞凋亡率降低,划痕愈合面积百分比增加,侵袭细胞数增多,cyclin D1、c-myc、β-catenin蛋白相对表达量升高(P<0.05);与纳武单抗+激动剂组比较,纳武单抗+激动剂+五味子素B组细胞存活率降低,细胞凋亡率升高,划痕愈合面积百分比减小,侵袭细胞数减少,cyclin D1、c-myc、β-catenin蛋白相对表达量降低(P<0.05)。结论:五味子素B能增强舌鳞癌细胞纳武单抗化疗敏感性,其可能机制是抑制Wnt/β-catenin信号通路。

Objective:To investigate the effect of schisandrin B on the sensitivity of human tongue squamous cell carcinoma to chemotherapy with nivolumab and the effect on secreted glycoprotein(Wnt)/β-catenin signaling pathway.Methods:Human tongue squamous cell carcinoma SCC15 cells in logarithmic growth stage were randomly divided into control group,nivolumab group,nivolumab+schisandrine B group,nivolumab+agonist group,and nivolumab+agonist+schisandrine B group.The cells in each group were treated with corresponding drug,then cell survival rate was detected by MTT assay,cell apoptosis rate was detected by flow cytometry,percentage of scar healing area was detected by scratch assay,number of invaded cells was detected by Transwell assay,and relative expression levels of cyclin D1,c-myc andβ-catenin protein were detected by Western blot.Results:Compared with the control group,the cell survival rate,the percentage of scratch healing area,the number of invasive cells and the relative expression levels of cyclin D1,c-myc andβ-catenin in nivolumab group were decreased(P<0.05).Compared with the nivolumab group,the cell survival rate reduced,apoptosis rate increased,the percentage of scratch healing area reduced,the number of invasive cells and the relative expression levels of cyclin D1,c-myc andβ-catenin were decreased in nivolumab+schisandrine B group(P<0.05),while the cell survival rate increased,the apoptosis rate decreased,the percentage of scratch healing area increased,and the number of invasive cells and the relative expression levels of cyclin D1,c-myc andβ-catenin in the nivolumab+agonist group were increased(P<0.05).Compared with nivolumab+schisandin B group,cell survival rate increased,apoptosis rate decreased,the percentage of scratch healing area,number of invasive cells and relative expression levels of cyclin D1,c-myc andβ-catenin in nivolumab+agonist+schisandin B group were increased(P<0.05).Compared with nivolumab+agonist group,the survival rate decreased,apoptosis rate increased,the percentage of scratch healing area,number of invasive cells and relative expression levels of cyclin D1,c-myc andβ-catenin in navuzumab+agonist+schisandin B group were decreased(P<0.05).Conclusion:Schisandrine B enhances the chemosensitivity of tongue squamous cell carcinoma cells to chemotherapy with nivolumab,and its possible mechanism is to inhibit Wnt/β-catenin signaling pathway.