Eg5小分子抑制剂SB-743921通过c-Myc/SP1/CDK1通路抑制去势抵抗性前列腺癌细胞恶性增殖的研究  

Eg5 inhibitor SB-743921 inhibits malignant biological features of castration-resistant prostate cancer cells through c-Myc/SP1/CDK1 signaling pathway

作　　者：徐礼臻 朱再生[1] 周鹏飞 刘全启[1] 徐旻 胡亮[1] XU Lizhen;ZHU Zaisheng;ZHOU Pengfei;LIU Quanqi;XU Min;HU Liang(Department of Urology,Affiliated Jinhua Hospital,Zhejiang University School of Medicine,Jinhua 321000,China)

机构地区：[1]浙江大学医学院附属金华医院泌尿外科,321000

出　　处：《浙江医学》2023年第15期1609-1614,1625,共7页Zhejiang Medical Journal

基　　金：金华市科学技术研究计划项目(2019-4-022)。

摘　　要：目的探讨驱动蛋白(Eg5)小分子抑制剂SB-743921对去势抵抗性前列腺癌(CRPC)细胞增殖、凋亡、细胞周期及迁移的影响以及相关分子机制。方法取对数生长期的人CRPC细胞株DU145,设置对照组、NC小干扰RNA(siRNA)组、Eg5 siRNA组、SB-743921(4 nmol/L)组、SB-743921(8 nmol/L)组。分别采用MTT法、流式细胞术、PI单染实验、划痕实验检测SB-743921对细胞增殖、凋亡、细胞周期及迁移的影响;进一步采用qRT-PCR、Western blot法检测SB-743921对细胞中c-Myc、SP1、周期蛋白依赖性激酶(CDK)1、E-钙黏蛋白(E-cadherin)、波形蛋白(Vimentin)mRNA及蛋白表达水平的影响。结果Eg5 siRNA组、SB-743921(4 nmol/L)组、SB-743921(8 nmol/L)组生长抑制率、细胞凋亡率、G_(2)/M期百分比均明显高于对照组(均P<0.05),而细胞迁移率均明显低于对照组(均P<0.05)。Eg5 siRNA组、SB-743921(4 nmol/L)组、SB-743921(8 nmol/L)组c-Myc、SP1、CDK1、Vimentin mRNA及蛋白表达水平均明显低于对照组(均P<0.05),E-cadherin mRNA及蛋白表达水平均明显高于对照组(均P<0.05);SB-743921(8 nmol/L)组CDK1、Vimentin蛋白表达水平均明显低于Eg5 siRNA组(均P<0.05)。结论SB-743921可通过c-Myc/SP1/CDK1信号通路阻断CRPC细胞的恶性增殖和迁移,诱导细胞凋亡并将细胞周期阻滞在G_(2)/M期。Objective To investigate the effects of kinesin-5(Eg5)inhibitor SB-743921 on the proliferation,apoptosis,cell cycle,invasion and metastasis of castration-resistant prostate cancer(CRPC)cells and the related mechanism.Methods Human CRPC DU145 cells were divided into control group,NC small interfering RNA(siRNA)group,Eg5 siRNA group,SB-743921(4 nmol/L)group and SB-743921(8 nmol/L)group.The effects of SB-743921 on cell proliferation,apoptosis,cell cycle,and migration were detected using MTT test,PI staining,flow cytometry and scratch assay,respectively.The expression of c-Myc,SP1,cyclin-dependent kinase 1(CDK1),E-cadherin,Vimentin mRNA and protein in DU145 cells were detected by qRT-PCR and Western blot,respectively.Results The growth inhibition rate,apoptosis rate,G2/M phase percentage of Eg5 siRNA group,SB-743921(4 nmol/L)group and SB-743921(8 nmol/L)group were significantly higher than those of the control group(all P<0.05),while cell migration rates were significantly lower than those of the control group(all P<0.05).The expression levels of c-Myc,SP1,CDK1,Vimentin mRNA and protein in the Eg5 siRNA group,SB-743921(4 nmol/L)group,and SB-743921(8 nmol/L)group were significantly lower than those of the control group(all P<0.05),while the expression levels of E-cadherin mRNA and protein were significantly higher than those of the control group(all P<0.05).The expression levels of CDK1 and Vimentin proteins in the SB-743921(8 nmol/L)group were significantly lower than those of the Eg5 siRNA group(all P<0.05).Conclusion SB-743921 can block the malignant proliferation and migration of CRPC cells through c-Myc/SP1/CDK1 signaling pathway,induce cell apoptosis and block cell cycle in G2/M phase.

关 键 词：驱动蛋白5 去势抵抗性前列腺癌 C-MYC SP1 周期蛋白依赖性激酶1 

分 类 号：R737.25[医药卫生—肿瘤]