摘要
目的:探讨芥子碱硫氰酸盐(sinapine thiocyanate,ST)对人胶质瘤T98细胞增殖、凋亡、迁移与侵袭,以及Wnt/β-连环蛋白(β-catenin)信号通路的影响。方法:以不同浓度(0、50、100和200μmol/L)的ST处理人胶质瘤T98细胞,分别采用CCK-8法和FCM法检测不同浓度的ST对T98细胞增殖率和凋亡率的影响。划痕愈合实验和Transwell小室法分别检测T98细胞的迁移和侵袭能力。计算机分子对接法分析ST与Wnt/β-catenin信号通路关键蛋白β-catenin的结合情况。最后采用蛋白质印迹法检测不同浓度ST处理后T98细胞中β-catenin蛋白的表达水平、凋亡相关蛋白Bcl-2与Bax的表达水平,以及基质金属蛋白酶2(matrix metalloproteinase-2,MMP-2)和MMP-9的表达水平。结果:不同浓度的ST能抑制T98细胞的增殖,并诱导细胞凋亡(P值均<0.05);ST还能明显抑制胶质瘤T98细胞的迁移和侵袭(P值均<0.05)。ST能与β-catenin的活性口袋稳定结合。ST可明显抑制T98细胞中β-catenin、Bcl-2、MMP-2和MMP-9蛋白的表达水平,而上调Bax蛋白的表达水平(P值均<0.05)。结论:ST可能通过结合Wnt/β-catenin信号通路关键分子β-catenin,抑制胶质瘤T98细胞的增殖、迁移和侵袭。
Objective:To investigate the effects of sinapine thiocyanate(ST)on the proliferation,migration and invasion of human glioma T98 cells,and on the Wnt/β-catenin signaling pathway.Methods:Human glioma T98 cells were treated with ST at different concentrations(0,50,100 and 200μmol/L),and then the relative cell proliferation rate was detected by CCK-8 method.The cell apoptotic rate was detected by flow cytometry.The cell migration and invasion abilities were detected by wound healing assay and Transwell chamber method.Computer molecular docking method was used to analyze the binding between ST andβ-catenin,a key protein of Wnt/β-catenin signaling pathway.Finally,the expressions of key proteinβ-catenin in Wnt/β-catenin signaling pathway,apoptosis-related BCL-2 and Bax proteins,as well as the matrix metalloproteinase(MMP)-2 and MMP-9 reflecting cytotoxicity in glioma T98 cells treated with different concentrations of ST were detected by Western blotting.Results:ST significantly inhibited the proliferation of glioma T98 cells and induced their apoptosis(both P<0.05).ST significantly inhibited the migration and invasion of glioma T98 cells(both P<0.05).ST stably binded to the active pocket ofβ-catenin.ST significantly reduced the expressions ofβ-catenin,Bcl-2,MMP-2 and MMP-9 proteins in T98 cells,while increased the expression of Bax protein(all P<0.05).Conclusion:ST may inhibit the proliferation,migration and invasion of glioma T98 cells by combiningβ-catenin,a key molecule in Wnt/β-catenin signaling pathway.
作者
谢明
曾智锐
陈鹏
周璐
朱俊德
XIE Ming;ZENG Zhirui;CHEN Peng;ZHOU Lu;ZHU Junde(Department of Human Anatomy,School of Basic Medical Sciences,Guizhou Medical University,Guian 550025,Guizhou Province,China;Department of Physiology,School of Basic Medical Sciences,Guizhou Medical University,Guian 550025,Guizhou Province,China;Key Laboratory of Pathogenesis and Drug Research of Common Chronic Diseases in Guizhou Province,Guian 550025,Guizhou Province,China;College of Clinical Medicine,Guizhou Medical University,Guizhou 550004,Guizhou Province,China)
出处
《肿瘤》
CAS
CSCD
北大核心
2020年第10期687-695,共9页
Tumor
基金
国家自然科学基金资助项目(编号:81660243)。
