To explore the effect of quercetin on the proliferation and apoptosis of HeLa cells, HeLa cells were incubated with quercetin at different concentrations. Cell viability was evaluated by MTT assay, cell apoptosis was ...To explore the effect of quercetin on the proliferation and apoptosis of HeLa cells, HeLa cells were incubated with quercetin at different concentrations. Cell viability was evaluated by MTT assay, cell apoptosis was detected by Annexin- V/PI double labeled cytometry and DNA ladder assay. Cell cycle was flow cytometrically determined and the morphological changes of the cells were ob- served under a fluorescence microscope after Hoechst 33258 staining and the apoptosis-related pro- teins in the HeLa cells were assessed by Western blotting. The results showed that quercetin signifi- cantly inhibited the growth of HeLa cells and induced obvious apoptosis in vitro in a time- and dose-dependent manner. Moreover, quercetin induced apoptosis of HeLa cells in cell cycle-dependent manner because quercetin could induce arrest of HeLa cells at G0/G1 phase. Quercetin treatment down-regulated the expression of the PI3K and p-Akt. In addition, quercetin could down-regulate ex- pression of bcl-2, up-regulate Bax, but exerted no effect on the overall expression of Akt. We are led to conclude that quercetin induces apoptosis via PI3k/Akt pathways, and quercetin has potential to be used as an anti-tumor agent against human cervix oancer.展开更多
目的研究蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用,并探讨其作用机制。方法采用流式细胞仪检测HeLa细胞的细胞周期分布相和细胞凋亡率;末端脱氧核苷酸转移酶(TdT)介导的脱氧核苷酸缺口末端标记(TdT-mediated dUTP nick end labeling)T...目的研究蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用,并探讨其作用机制。方法采用流式细胞仪检测HeLa细胞的细胞周期分布相和细胞凋亡率;末端脱氧核苷酸转移酶(TdT)介导的脱氧核苷酸缺口末端标记(TdT-mediated dUTP nick end labeling)TUNEL法检测凋亡细胞;通过免疫细胞化学SP法测定凋亡相关基因产物p53、Fas、bcl-2、Bax蛋白的表达情况。结果(1)蛴螬粗提物可以诱导HeLa细胞发生凋亡,加药组出现亚二倍体峰,并且G0/G1期细胞比例显著增加(P<0.01),而S期和G2/M期细胞比例则明显下降,将细胞阻滞在G0/G1期;(2)蛴螬粗提物作用细胞后凋亡细胞数目随时间延长而增多,凋亡指数与药物处理呈时间依赖性和剂量依赖性;(3)蛴螬粗提物作用后bcl-2、p53蛋白随提取物浓度表达均下降,Bax、Fas蛋白表达上升,四种蛋白表达各组间比较差异均有具有统计学意义(P<0.01)。结论(1)蛴螬粗提物对人宫颈癌HeLa细胞具有诱导凋亡作用;(2)蛴螬粗提取物诱导凋亡作用机制可能与细胞周期发生G0/G1期阻滞有关;并且通过下调bcl-2、p53蛋白表达,上调Bax、Fas蛋白表达,经由细胞凋亡的死亡受体通路和线粒体通路完成凋亡的启动和执行。展开更多
基金supported by the National Natural Science Foundation of China(No.81071663)
文摘To explore the effect of quercetin on the proliferation and apoptosis of HeLa cells, HeLa cells were incubated with quercetin at different concentrations. Cell viability was evaluated by MTT assay, cell apoptosis was detected by Annexin- V/PI double labeled cytometry and DNA ladder assay. Cell cycle was flow cytometrically determined and the morphological changes of the cells were ob- served under a fluorescence microscope after Hoechst 33258 staining and the apoptosis-related pro- teins in the HeLa cells were assessed by Western blotting. The results showed that quercetin signifi- cantly inhibited the growth of HeLa cells and induced obvious apoptosis in vitro in a time- and dose-dependent manner. Moreover, quercetin induced apoptosis of HeLa cells in cell cycle-dependent manner because quercetin could induce arrest of HeLa cells at G0/G1 phase. Quercetin treatment down-regulated the expression of the PI3K and p-Akt. In addition, quercetin could down-regulate ex- pression of bcl-2, up-regulate Bax, but exerted no effect on the overall expression of Akt. We are led to conclude that quercetin induces apoptosis via PI3k/Akt pathways, and quercetin has potential to be used as an anti-tumor agent against human cervix oancer.
文摘目的研究蛴螬粗提物对人宫颈癌HeLa细胞诱导凋亡作用,并探讨其作用机制。方法采用流式细胞仪检测HeLa细胞的细胞周期分布相和细胞凋亡率;末端脱氧核苷酸转移酶(TdT)介导的脱氧核苷酸缺口末端标记(TdT-mediated dUTP nick end labeling)TUNEL法检测凋亡细胞;通过免疫细胞化学SP法测定凋亡相关基因产物p53、Fas、bcl-2、Bax蛋白的表达情况。结果(1)蛴螬粗提物可以诱导HeLa细胞发生凋亡,加药组出现亚二倍体峰,并且G0/G1期细胞比例显著增加(P<0.01),而S期和G2/M期细胞比例则明显下降,将细胞阻滞在G0/G1期;(2)蛴螬粗提物作用细胞后凋亡细胞数目随时间延长而增多,凋亡指数与药物处理呈时间依赖性和剂量依赖性;(3)蛴螬粗提物作用后bcl-2、p53蛋白随提取物浓度表达均下降,Bax、Fas蛋白表达上升,四种蛋白表达各组间比较差异均有具有统计学意义(P<0.01)。结论(1)蛴螬粗提物对人宫颈癌HeLa细胞具有诱导凋亡作用;(2)蛴螬粗提取物诱导凋亡作用机制可能与细胞周期发生G0/G1期阻滞有关;并且通过下调bcl-2、p53蛋白表达,上调Bax、Fas蛋白表达,经由细胞凋亡的死亡受体通路和线粒体通路完成凋亡的启动和执行。
