槲皮素诱导HeLa细胞凋亡及caspase-3、caspase-8活化对凋亡影响的研究

顾超 徐水凌 唐文稳 朱逢佳 徐倩 高帆

中国药学杂志 ›› 2011, Vol. 46 ›› Issue (8) : 595-599.

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PDF(740 KB)
中国药学杂志 ›› 2011, Vol. 46 ›› Issue (8) : 595-599.
论著

槲皮素诱导HeLa细胞凋亡及caspase-3、caspase-8活化对凋亡影响的研究

  • 顾超,徐水凌*,唐文稳,朱逢佳,徐倩,高帆
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HeLa Cell Apoptosis Induced by Quercetin and Effects of Caspase-3, Caspase-8 Activation on Apoptosis

  • GU Chao, XU Shui-ling*, TANG Wen-wen, ZHU Feng-jia, XU Qian, GAO Fan
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摘要

目的 研究槲皮素(quercetin诱导HeLa细胞(人宫颈癌细胞凋亡的作用,以及caspase-3、caspase-8活化对凋亡的影响。方法 采用噻唑蓝(MTT法观察槲皮素对HeLa细胞(人宫颈癌细胞和L-02细胞(正常人肝细胞的生长抑制作用;DAPI染色后荧光显微镜观察细胞的形态学变化;通过DNA琼脂糖凝胶电泳进行DNA片段化分析,FITC-Annexin V/PI荧光标记流式细胞术检测细胞凋亡情况;比色法测定槲皮素诱导HeLa细胞凋亡过程中caspase-3、caspase-8活性变化。结果 MTT结果显示,槲皮素可选择性抑制HeLa细胞的生长,并呈剂量、时间依赖性,而正常人肝细胞L-02不出现生长抑制(P>0.05。160.0 μmol·L-1槲皮素处理HeLa细胞72 h,DAPI染色也可见核浓缩及边缘现象,DNA电泳出现特征性的凋亡条带。160.0 μmol·L-1槲皮素处理HeLa细胞24、48和72 h的凋亡率分别为(9.4±2.1%、(30.1±6.5%和(59.8±9.5%,caspase-3活性在48 h达最高(1.68±0.07U·μg -1,而caspase-8活性则在24 h达最高(1.83±0.06U·μg-1,两者与对照组相比均具有显著性差异(P<0.05。结论 槲皮素可特异性地诱导人HeLa细胞凋亡,其诱导凋亡的机制可能与caspase-3、caspase-8活化有关。

Abstract

OBJECTIVE To study the apoptosis of HeLa cells(human cervical cancer cells induced by quercetin and the effects of caspase-3,caspase-8 activation on the apoptosis. METHODS Colorimetric MTT assay was used to measure the inhibition of quercetin in HeLa cells and L-02 cells. The morphological changes of HeLa cells were observed by fluorescence microscope after DAPI staining. The apoptosis of treated cells were examined by DNA agarose gel electrophoresis analysis of DNA fragmentation and flow cytometry using fluorescein labeling FITC-Annexin V/PI, respectively. The activity of caspase-3 and caspase-8 in apoptosis as detected by colorimetric assay. RESULTS MTT results showed that quercetin could inhibit the growth of HeLa cells in dose-and time-dependent manner significantly, but couldnt inhibit the growth of L-02 cells(P>0.05.The phenomenon of nuclear condensation and marginalization by DAPI staining and the characteristic bands of apoptosis by DNA electrophoresis were found in HeLa cells treated with quercetin at 160.0 μmol·L-1for 72 h. The rates of apoptosis of HeLa cells which were treated with quercetin at 160.0 μmol·L-1for 24, 48 and 72 h were(9.4±2.1%、(30.1±6.5% and(59.8±9.5%, respectively. The maximal activities of intracellular caspase-3 at 48 h and caspazse-8 at 24 h were(1.68±0.07 and (1.83±0.06U·μg-1, respectively. There was significantly difference compared with the control group(P<0.05.CONCLUSION Quercetin can specifically induce the apoptosis of HeLa cells, and intracellular caspase-3 and caspase-8 are closely associated with the apoptosis.

关键词

槲皮素 / HeLa细胞 / 细胞凋亡 / 半胱氨酸天冬氨酸蛋白酶

Key words

quercetin / HeLa cell lines / apoptosis

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顾超 徐水凌 唐文稳 朱逢佳 徐倩 高帆. 槲皮素诱导HeLa细胞凋亡及caspase-3、caspase-8活化对凋亡影响的研究[J]. 中国药学杂志, 2011, 46(8): 595-599
GU Cho;XU Shui-ling;TNG Wen-wen;ZHU Feng-ji;XU Qin;GO Fn. HeLa Cell Apoptosis Induced by Quercetin and Effects of Caspase-3, Caspase-8 Activation on Apoptosis[J]. Chinese Pharmaceutical Journal, 2011, 46(8): 595-599
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