摘要
目的:探讨水苏碱(STA)通过调节Yes相关蛋白(YAP)/具有PDZ结合基序的转录辅激活因子(TAZ)信号通路对膀胱癌细胞恶性生物学行为和糖酵解的影响。方法:以膀胱癌T24细胞为研究对象,将T24细胞分为对照组(不加药物处理),STA低(STA-L)、中(STA-M)、高剂量(STA-H)组,YAP/TAZ信号通路激活剂(GA-017)组和GA-017+STA-H组。CCK-8法检测细胞增殖活性;克隆形成实验检测细胞克隆形成能力;Transwell实验测定细胞侵袭能力;流式细胞术检测细胞凋亡;利用试剂盒检测影响糖酵解的因子水平(ATP、乳酸生成量、葡萄糖消耗量);免疫印迹检测caspase-3、Bcl-2、MMP-9、YAP、TAZ蛋白表达。结果:与对照组比较,STA各组及GA-017+STA-H组T24细胞的增殖活性、克隆形成率、侵袭数目、MMP-9蛋白表达、Bcl-2蛋白表达、ATP水平、乳酸生成量、葡萄糖消耗量显著下降,细胞凋亡率及caspase-3表达显著上升,STA-M、STA-H组及GA-017+STA-H的YAP、TAZ蛋白表达下降,差异具有统计学意义;GA-017组上述指标与STA-H组呈现相反的变化趋势;与STA-H组比较,GA-017+STA-H组可在一定程度逆转STA对于T24细胞恶性生物学和糖酵解行为的抑制作用。结论:STA可通过抑制YAP/TAZ信号通路表达,从而抑制膀胱癌细胞T24的增殖、侵袭、糖酵解,促进T24细胞凋亡。
Objective:To investigate the influence of stachydrine(STA)on the malignant biological behavior and glycolysis of bladder cancer cells by regulating Yes-associated protein(YAP)/transcriptional coactivator with PDZbinding motif(TAZ)signaling pathway.Methods:Bladder cancer T24 cells,taken as the research subject,were divided into control group(without drug treatment),low dose(STA-L),medium dose(STA-M),high dose(STA-H)STA groups,YAP/TAZ signaling pathway activator(GA-017)group,and GA-017+STA-H group.Cell proliferation activity was detected by CCK-8 method.Clonogenic assay was used to detect the clonogenic capacity of cells.Transwell assay was used to measure the invasive ability of cells.Cell apoptosis was detected by flow cytometry.The levels of factors affecting glycolysis(ATP,lactic acid production,glucose consumption)were detected with the kits,and the protein expressions of caspase-3,Bcl-2,MMP-9,YAP,and TAZ were detected by Western blotting.Results:Compared with the control group,the proliferation activity,colony formation rate,invasion number,MMP-9 protein expression,Bcl-2 protein expression,ATP level,lactic acid production,and glucose consumption of T24 cells in the three STA groups and the GA-017+STA-H group were decreased obviously,the apoptosis rate and caspase-3 expression were increased obviously.The expression levels of YAP and TAZ proteins in the STA-M,STA-H groups and the GA-017+STA-H group were decreased obviously;and the above indexes in the GA-017 group showed an opposite trend to those in the STA-H group.Compared with the STA-H group,the GA-017+STA-H group could reverse the inhibition of STA on the malignant biology and glycolysis of T24 cells to some extent.Conclusion:STA can inhibit the proliferation,invasion,glycolysis of bladder cancer cell T24,and promote the apoptosis of T24 cells by inhibiting the expression of YAP/TAZ signaling pathway.
作者
袁俊峰
林波
陈侃
王书龙
Yuan Junfeng;Lin Bo;Chen Kan;Wang Shulong(Department of Urology,People's Armed Police Force Hospital of Hubei Province,Wuhan 430070;Department of Urology,General Hospital of the Central Theater Command of the People's Liberation Army,Wuhan 430070,China)
出处
《解剖学杂志》
CAS
2024年第4期308-313,共6页
Chinese Journal of Anatomy
关键词
水苏碱
膀胱癌
增殖
侵袭
糖酵解
YAP/TAZ信号通路
stachydrine
bladder cancer
proliferation
invasion
glycolysis
YAP/TAZ signaling pathway
