摘要
目的:探究重楼皂苷Ⅰ介导Wntβ-catenin信号通路对鼻咽癌CNE1细胞生长和上皮间质转化的机制。方法:体外培养鼻咽癌CNE1细胞,并分为空白组(Control)、低浓度重楼皂苷Ⅰ组(5μmol/L)、中浓度重楼皂苷Ⅰ组(10μmol/L)、高浓度重楼皂苷Ⅰ组(20μmol/L),分别使用对应剂量的重楼皂苷Ⅰ预处理。使用brdu实验检测细胞生长情况;使用Hoechst 33258染色检测细胞凋亡情况;使用Transwell检测细胞侵袭情况;使用蛋白质印记法检测N-cadherin、E-cadherin、Wnt、β-catenin、TCF4表达情况;使用免疫荧光检测Vimentin表达情况。结果:与空白组比较,使用重楼皂苷Ⅰ处理的各组鼻咽癌CNE1细胞凋亡率、E-cadherin蛋白表达水平显著升高,且随着使用重楼皂苷Ⅰ的浓度的升高上述指标显著升高(P<0.05);与空白组比较,使用重楼皂苷Ⅰ处理的各组鼻咽癌CNE1细胞Brdu染色单位面积内阳性率、单位面积内鼻咽癌CNE1细胞侵袭细胞数目、N-cadherin,Wnt,β-catenin及TCF4蛋白表达水平、每个细胞内免疫荧光检测Vimentin发光点数显著降低,且随着使用重楼皂苷Ⅰ的浓度的升高上述指标降低的越显著(P<0.05)。结论:重楼皂苷Ⅰ可以通过介导Wntβ-catenin信号通路抑制鼻咽癌CNE1细胞上皮间质转化实现抑制其侵袭、迁移等恶性生物学行为,并促进鼻咽癌CNE1细胞的凋亡且在一定浓度范围内呈浓度依赖性。
Objective:To investigate the mechanism of polyphyllin Ⅰ(PP Ⅰmediating Wntβ-catenin signaling pathway on growth of nasopharyngeal carcinoma(NPC)CNE1 cells and epithelial-mesenchymal transition.Methods:NPC CNE1 cells were cultured in vitro.They were divided into blank group(Control),low-concentration PP Ⅰ group(5 μmol/L),medium-concentration PP Ⅰ group(10 μmol/L)and high-concentration PP Ⅰ group(20 μmol/L),given corresponding dose of PP Ⅰ,respectively.Cell growth was detected by brdu assay.The apoptosis was detected by Hoechst 33258 staining.Cell invasion was detected by Transwell.The expression of N-cadherin,E-cadherin,Wnt,β-catenin and TCF4 was detected by Western blot.The immunofluorescence was performed to detect Vimentin expression.Results:Compared with the blank group,the apoptosis rates of NPC CNE1 cells and expression levels of E-cadherin protein in each group treated with PP Ⅰ were significantly increased.The above indexes were significantly increased with the increase of PP Ⅰ concentration(P<0.05).Compared with the blank group,the positive rates of NPC CNE1 cells by Brdu staining per unit area,number of invasive NPC CNE1 cells per unit area,expression levels of N-cadherin,Wnt,β-catenin and TCF4 protein and number of Vimentin luminescence points in each cell detected by immunofluorescence in each group treated with PPI were significantly decreased.The decrease of the above-mentioned indexes was more significant with increase of PP Ⅰ concentration(P<0.05).Conclusion:PP Ⅰ can inhibit NPC CNE1 cells epithelial-mesenchymal transition by mediating Wntβ-catenin signaling pathway thus inhibiting malignant biological behavior such as invasion and migration,and promote apoptosis of NPC CNE1 cells,showing concentration-dependence within a certain concentration range.
作者
王波涛
孙斌
祝康
夏翠
高天喜
WANG Botao;SUN Bin;ZHU Kang;XIA Cui;GAO Tianxi(Department of Otolaryngology head and neck surgery,second affiliated Hospital of Xi′an Jiaotong University,Xi′an 710004,China)
出处
《世界中医药》
CAS
2020年第24期3782-3786,3791,共6页
World Chinese Medicine
基金
陕西省科学技术厅一般项目(2017SF-210)。
