摘要
目的探讨组蛋白乙酰转移酶KAT7抑制剂WM-3835对膀胱癌细胞增殖和迁移能力的影响以及可能的机制。方法用不同浓度的WM-3835(0、10、20、30、40μmol/L)处理人输尿管上皮永生化细胞系SV-HUC-1,膀胱癌细胞系UM-UC-3和T24,48 h后通过噻唑蓝实验观察WM-3835对正常尿路上皮细胞和膀胱癌细胞增殖能力的影响;采用流式细胞术分析WM-3835对膀胱癌细胞周期分布的影响;通过划痕实验和Transwell实验检测WM-3835对膀胱癌细胞迁移能力的影响;通过实时荧光定量PCR分析和免疫印迹实验检测增殖标志物细胞周期蛋白D1(cyclin-D1)、增殖细胞核抗原(PCNA)、基质金属蛋白酶9(MMP9)以及神经钙粘素(N-cadherin)的mRNA和蛋白表达水平的变化。结果WM-3835可以显著抑制膀胱癌细胞增殖,同一浓度下对正常尿路上皮细胞杀伤作用较小;不同浓度的WM-3835处理膀胱癌细胞UM-UC-3和T24后,膀胱癌细胞周期出现G0/G1期阻滞,细胞的增殖能力受到有效抑制,迁移能力显著减弱。实时荧光定量PCR分析和免疫印迹实验显示,膀胱癌细胞经WM-3835处理后,增殖相关标志物cyclin-D1和PCNA以及上皮间质转化相关标志物MMP9和N-cadherin的mRNA及蛋白表达水平均出现下调。结论WM-3835能够抑制膀胱癌细胞的增殖和迁移能力,有成为膀胱癌化疗用药的潜力。
Objective To investigate the effects of WM-3835,a histone acetyltransferase KAT7(KAT7)inhibitor,on the proliferation and migration of bladder cancer cells and to explore the possible mechanism.Methods Human ureteral epithelial immortalized cell line SV-HUC-1,and bladder cancer cell lines UM-UC-3 and T24 were treated with different concentrations of WM-3835(0,10,20,30,40μmol/L).After 48 hours,the effects of WM-3835 on the proliferation,cell cycle distribution and migration of cells were detected with MTT assay,flow cytometry,scratch and Transwell assay,respectively.The expressions of cyclin D1(cyclin D1),proliferating nuclear antigen(PCNA),matrix metalloproteinase 9(MMP9)and neurocadherin(N-cadherin)were detected with Western blotting and real-time quantitative PCR.Results WM-3835 significantly inhibited the proli-feration of bladder cancer cells in a dose-dependent manner.After treatment with WM-3835,the cycle of UM-UC-3 and T24 cells were blocked in the G0/G1 phase,the proliferation was effectively inhibited,and the migration was significantly wea-kened.The expressions of cyclin-D1,PCNA,MMP9 and N-cadherin were down-regulated.Conclusion WM-3835 can inhibit the proliferation and migration of bladder cancer cells,and has the potential as a chemotherapeutic agent for bladder cancer.
作者
王旺
熊康平
钱开宇
王行环
WANG Wang;XIONG Kangping;QIAN Kaiyu;WANG Xinghuan(Department of Urology,Zhongnan Hospital of Wuhan University,Wuhan 430071;Hubei Provincial Human Genetic Resources Conservation Center,Zhongnan Hospital of Wuhan University,Wuhan 430071,China)
出处
《现代泌尿外科杂志》
CAS
2023年第6期500-505,共6页
Journal of Modern Urology
基金
湖北省科技厅科技服务人才项目(No.2022EJD001)。