摘要
目的:探讨法舒地尔(FSDE)调节RhoA/ROCK信号通路对膀胱癌细胞增殖、凋亡和血管生成的影响。方法:将膀胱癌T24细胞分为对照组,FSDE低、中、高剂量组,FSDE高剂量+Naciclasine(RhoA激活剂)组。MTT和EdU检测细胞增殖;Transwell实验检测细胞迁移和侵袭;流式细胞术检测细胞凋亡率;Matrigel胶三维培养观察血管生成情况;免疫印迹检测细胞血管内皮生长因子A(VEGF-A)、血管内皮钙黏蛋白(VE-cadherin)、Ki-67、基质金属蛋白酶9(MMP-9)、caspase-3、RhoA、ROCK蛋白表达水平。结果:对照组细胞具有良好的管腔结构;与对照组相比,FSDE低、中、高剂量组T24细胞管腔结构有不同程度损坏,OD490(24 h、48 h)值和细胞增殖率、细胞迁移和侵袭个数、VEGF-A、VE-cadherin、Ki-67、MMP-9、RhoA、ROCK蛋白表达显著降低,细胞凋亡率、caspase-3蛋白表达显著升高;Naciclasine可部分逆转FSDE对T24细胞的抑制作用。结论:FSDE可抑制膀胱癌细胞恶性生物学行为,可能是通过阻断RhoA/ROCK信号通路激活实现。
Objective:To investigate the effect of fasudil on the proliferation,apoptosis and angiogenesis of bladder cancer cells by regulating RhoA/ROCK signaling pathway.Methods:T24 cells of bladder cancer were divided into control group,low-dose fasudil group,medium-dose fasudil group,high-dose fasudil group,and high-dose fasudil+Naciclassine(RhoA activator)group.MTT and EdU assays were adopted to detect cell proliferation.Transwell assay was applied to detect cell migration and invasion.Flow cytometry was employed to detect cell apoptosis rate.Threedimensional Matrigel culture was performed to observe angiogenesis.Western blotting was utilized to detect the expression levels of vascular endothelial growth factor A(VEGF-A),vascular endothelial cadherin(VE cadherin),Ki-67,MMP-9,caspase-3,RhoA,and ROCK proteins in cells.Results:The control group cells had good lumen structure.Compared with the control group,the lumen structure of T24 cells in the low,medium,and high-dose fasudil groups showed varying degrees of damage,and the OD490 values(at 24 h and 48 h),cell viability,numbers of cell migration and invasion,the expression of VEGF-A,VE cadherin,Ki-67,MMP-9,RhoA,and ROCK proteins were obviously reduced,while the apoptosis rate and the expression of caspase-3 protein were obviously increased.Naciclassine partially reversed the inhibitory effect of fasudil on T24 cells.Conclusion:Fasudil can inhibit the malignant biological behavior of bladder cancer cells,possibly by blocking the RhoA/ROCK signaling pathway activation.
作者
陈蕾
何超芹
陈伟前
Chen Lei;He Chaoqin;Chen Weiqian(Department of Pharmacy,Jinhua Municipal Central Hospital,Jinhua 321000,China)
出处
《解剖学杂志》
2025年第3期235-240,共6页
Chinese Journal of Anatomy
作者简介
第一作者:陈蕾,E-mail:liem7978662@163.com。
