摘要
目的探索红景天苷(sali)对人肺腺癌细胞的上皮-间质转化的影响及其作用机制。方法将肺腺癌PC9细胞随机分为6组:对照组(Con),转化生长因子β1(TGF-β1)组,sali 1μg·mL^(-1)组,sali 5μg·mL^(-1)组,sali 10μg·mL^(-1)组,sali 20μg·mL^(-1)组。肺腺癌PC9细胞在5 ng·mL^(-1)的TGF-β1刺激30 min后,再各组给予对应浓度sali干预,Con组不做TGF-β1和sali干预处理。各组分别在干预后24,48,72 h后进行MTT检测,绘制生长曲线;在各时间段显微镜下观察细胞形态,以流式细胞仪检测细胞凋亡率,以Western blotting检测上皮细胞钙粘蛋白(E-cadherin)、神经型钙粘蛋白(N-cadherin)、波形蛋白(Vimentin)、平滑肌肌动蛋白-α(α-SMA)、纤维连接蛋白(FN),β-连环蛋白(β-catenin)的表达。结果与TGF-β1组比较,sali 5μg·mL^(-1)组,sali 10μg·mL^(-1)组和sali 20μg·mL^(-1)组细胞增殖力均显著降低(P<0.05)。Con组细胞形态多成椭圆形或者圆形,TGF-β1组大部分细胞形态呈梭形,相邻细胞间连接也变得疏松,而sali 5μg·mL^(-1)组,sali 10μg·mL^(-1)组和sali 20μg·mL^(-1)组细胞形态均多半呈椭圆形,细胞之间连接紧密。sali 20μg·mL^(-1)组的细胞凋亡率均显著高于sali 5μg·mL^(-1)组和sali 1μg·mL^(-1)组;和TGF-β1组比较,sali 1μg·mL^(-1)组的标记蛋白无显著差异(P>0.05);sali 5μg·mL^(-1)组,sali 10μg·mL^(-1)组和sali 20μg·mL^(-1)组的N-cadherin,FN,Vimentin,α-SMA的表达均显著降低,而E-cadherin,β-catenin的表达显著升高(P<0.05)。结论TGF-β1可诱导肺癌上皮细胞向间质细胞转变;红景天苷可能通过上调β-catenin抑制肺癌细胞中上皮-间质转化(EMT)的发生。
Objective To explore the effect and its mechanism of salidroside on epithelial mesenchymal transition in lung adenocarcinoma cells.Methods PC9 lung adenocarcinoma cells were randomly divided into 6 groups:control group,TGF-βgroup,Sali 1μg·mL^(-1)group,Sali 5μg·mL^(-1)group,Sali 10μg·mL^(-1)group,Sali 20μg·mL^(-1)group.PC9 cells was treated with transforming growth factorβ1(TGF-β1)at 5 ng·mL^(-1)for 30 min.Then each group was intervened with salidroside of corresponding concentration for 24,48,72 h,respectively,and control group was not treated with TGF-β1 and salidroside.After the intervention,MTT test was performed at 24,48 and 72 hours after intervention in each group,and growth curve was drawn;the cell morphology was observed under the microscope,and the apoptosis rate was detected by flow cytometry.Western blotting was used to detect E-cadherin,N-cadherin,vimentin,smooth muscle actin-α(α-SMA),fibronectin(FN),andβ-Catenin expression.Results The cell proliferations of Sali 5μg·mL^(-1)group,Sali 10μg·mL^(-1)and Sali 20μg·mL^(-1)group were significantly decreased(P<0.05).Cell shape in the control group was oval or round,but most cell morphology in TGF-β1 group was spindle and connections between adjacent cells became loose.And cell morphology in Sali 5μg·mL^(-1)group,Sali 10μg·mL^(-1)group,and Sali 20μg·mL^(-1)group were mostly oval with tight connections between the cells.The apoptosis rate of Sali 20μg·mL^(-1)group was significantly higher than that in Sali 5μg·mL^(-1)group and Sali 1μg·mL^(-1)group.Compared with TGF-β1 group,there was no significant difference between Sali 1 g·mL^(-1)group(P>0.05);β-Catenin expression in Sali 5μg·mL^(-1)group was significantly increased(P<0.05).Conclusion TGF-β1 can induce epithelial cells to mesenchymal cells in lung cancer cell;salidroside may inhibit EMT in lung cancer cells through the upregulation ofβ-catenin.
作者
张鹏
孙婉瑾
李嫚
徐玉婷
ZHANG Peng;SUN Wanjin;LI Man;XU Yuting(Department of Pharmaceutical Affairs,Hubei Provincial Hospital of Traditional Chinese Medicine,Wuhan 430074,China;Department of Integrated TCM&Western Medicine,Hubei Cancer Hospital,Wuhan 430079,China)
出处
《医药导报》
CAS
北大核心
2021年第8期1005-1010,共6页
Herald of Medicine
基金
中国中医科学院中医基础理论研究所协同创新科研专项基金资助项目(YZ-1850)
湖北省自然科学基金资助项目(2011CDC031)。
关键词
红景天苷
Β-CATENIN
人肺腺癌细胞
上皮-间质转化
Salidroside
β-catenin
Human lung adenocarcinoma cells
Epithelial mesenchymal transition
Vimentin
