CAJ | 학술논문

目的研究自噬在槲皮素诱导的人膀胱癌BIU-87细胞凋亡中的作用.方法以不同浓度的槲皮素处理人膀胱癌BIU-.87细胞,采用四甲基偶氮唑蓝(MTT)法检测槲皮素对BIU-87细胞增殖的抑制作用,采用单丹磺酰尸胺(MDC)荧光染色、Hochest 33258染色分别观察槲皮素处理后BIU-87细胞的自噬和凋亡现象,采用流式细胞仪检测阻断自噬对BIU-87细胞凋亡的影响.以乳酸脱氢酶(LDH)检测试剂盒检测3-甲基腺嘌呤阻断自噬后对槲皮素细胞毒作用的影响.结果槲皮素对BIU-87细胞的增殖有明显的抑制作用,且此抑制作用呈明显的时间和浓度依赖性,200 μmol/L槲皮素作用72 h时对BIU-87细胞的增殖抑制率达89.2％.槲皮素可以诱导BIU-87细胞发生自噬和凋亡,并且自噬先于凋亡出现；阻断自噬后可使BIU-87细胞的凋亡峰明显增高.在槲皮素处理前后,分别以3-甲基腺嘌呤阻断自噬,可观察到槲皮素对BIU-87细胞的细胞毒作用均增强.结论槲皮素能明显抑制BIU-87细胞的增殖,并在诱导细胞凋亡之前先诱导自噬发生.在槲皮素诱导BIU-87细胞凋亡的过程中,自噬在早期能够保护BIU-87细胞,延迟凋亡,减少因槲皮素导致的细胞死亡.
목적 연구자서재곡피소유도적인방광암BIU-87세포조망중적작용.방법 이불동농도적곡피소처리인방광암BIU-.87세포,채용사갑기우담서람(MTT)법검측곡피소대BIU-87세포증식적억제작용,채용단단광선시알(MDC)형광염색、Hochest 33258염색분별관찰곡피소처리후BIU-87세포적자서화조망현상,채용류식세포의검측조단자서대BIU-87세포조망적영향.이유산탈경매(LDH)검측시제합검측3-갑기선표령조단자서후대곡피소세포독작용적영향.결과 곡피소대BIU-87세포적증식유명현적억제작용,차차억제작용정명현적시간화농도의뢰성,200 μmol/L곡피소작용72 h시대BIU-87세포적증식억제솔체89.2％.곡피소가이유도BIU-87세포발생자서화조망,병차자서선우조망출현；조단자서후가사BIU-87세포적조망봉명현증고.재곡피소처리전후,분별이3-갑기선표령조단자서,가관찰도곡피소대BIU-87세포적세포독작용균증강.결론 곡피소능명현억제BIU-87세포적증식,병재유도세포조망지전선유도자서발생.재곡피소유도BIU-87세포조망적과정중,자서재조기능구보호BIU-87세포,연지조망,감소인곡피소도치적세포사망.
Objective To explore the role of autophagy in quercetin (Que)-induced apoptosis in human bladder carcinoma BIU-87 cells in vitro.Methods 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.Results 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.Conclusions 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.