Wei Liang, Liu Jian-jun, Cao Jun, Du Ning-chao, Ji Li-na, Yang Xiao-liang
Department of Urological Surgery, Affiliated Hospital of Guangdong Medical College, Zhanjiang 524023, China.
Zhonghua Zhong Liu Za Zhi. 2012 Jun;34(6):414-8. doi: 10. 3760/cma.j.issn.0253-3766.2012.06.004.
To explore the role of autophagy in quercetin (Que)-induced apoptosis in human bladder carcinoma BIU-87 cells in vitro.
To determine the proliferative inhibition by MTT colorimetric assay after treating BIU-87 cells with quercetin at various concentrations. To identify autophagy and apoptosis in the BIU-87 cells after Que treatment by monodansylcadaverin (MDC) and Hoechst 33258 fluorescent staining, respectively. To examine the cytotoxic effect of Que and influence of autophagy on apoptosis by studying LDH leakage rate and flow cytometry, after blocking the autophagy with 3-methlyadenine (3-MA), a specific autophagy inhibitor.
There was an obvious inhibitory effect of Que on the proliferation of BIU-87 cells in a time- and dose-dependent manner. The inhibition rate of BIU-87 cells after 200 µmol/L Que treatment for 72 hours was 89.2%. Autophagy and apoptosis were induced and detected in Que-treated BIU-87 cells and autophagy occurred earlier than apoptosis. The apoptosis peak became much higher after the autophagy was blocked. Whenever the autophagy was blocked before or after Que treatment, the Que-induced cytotoxicity in BIU-87 cells was enhanced.
Quercetin significantly inhibits the proliferation of BIU-87 cells, and the autophagy is induced earlier than apoptosis. In the process of Que-induced apoptosis of BIU-87 cells, autophagy may play a protective role at the initiation phase, delay apoptosis and reduce the Que-induced death of BIU-87 cells.
探讨自噬在槲皮素(Que)诱导人膀胱癌BIU-87细胞体外凋亡中的作用。
用不同浓度的槲皮素处理BIU-87细胞后,采用MTT比色法测定其增殖抑制率。分别用单丹磺酰尸胺(MDC)和Hoechst 33258荧光染色鉴定Que处理后BIU-87细胞中的自噬和凋亡。在用特异性自噬抑制剂3-甲基腺嘌呤(3-MA)阻断自噬后,通过研究乳酸脱氢酶(LDH)泄漏率和流式细胞术,检测Que的细胞毒性作用以及自噬对凋亡的影响。
Que对BIU-87细胞的增殖有明显的抑制作用,呈时间和剂量依赖性。200μmol/L Que处理72小时后,BIU-87细胞的抑制率为89.2%。在Que处理的BIU-87细胞中诱导并检测到自噬和凋亡,且自噬发生早于凋亡。自噬被阻断后,凋亡峰明显升高。无论在Que处理前还是处理后阻断自噬,Que诱导的BIU-87细胞毒性均增强。
槲皮素显著抑制BIU-87细胞的增殖,且自噬诱导早于凋亡。在Que诱导BIU-87细胞凋亡的过程中,自噬可能在起始阶段起保护作用,延迟凋亡并减少Que诱导的BIU-87细胞死亡。