Diallyl sulfide induces cell cycle arrest and apoptosis in HeLa human cervical cancer cells through the p53, caspase- and mitochondria-dependent pathways
Diallyl sulfide induces cell cycle arrest and apoptosis in HeLa human cervical cancer cells through the p53, caspase- and mitochondria-dependent pathways
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DOI：
10.3892/ijo.2011.973
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发表时间：
2011-06-01
影响因子：
5.2
通讯作者：
Chung, Jing-Gung
中科院分区：
文献类型：
--
作者：
Wu, Ping-Ping;Chung, Hui-Wen;Chung, Jing-Gung
二烯丙基硫醚（DAS），大蒜的主要活性成分之一，在体外引起癌细胞的生长抑制，并在实验环境中促进体内免疫反应。但其对人宫颈癌细胞周期和凋亡的诱导作用尚不清楚。本研究旨在探讨DAS在体外对人宫颈癌细胞的抗肿瘤作用及其机制。采用形态学观察、细胞活力测定、DAPI染色、彗星试验、Western blotting和共聚焦显微镜观察等方法，研究了不同浓度的药物对人宫颈癌细胞系HeLa的细胞毒作用和细胞凋亡的影响。结果表明，DAS处理24-72 h导致细胞活力明显下降，并呈时间和剂量依赖性。流式细胞仪分析表明，75 μ MDAS处理48小时诱导HeLa细胞G 0/G1期阻滞和亚G1期（凋亡）。用DAPI染色，在荧光显微镜下观察到DAS暴露后的HeLa细胞中典型的凋亡细胞核改变。用不同浓度的DAS处理的细胞也显示出典型的凋亡变化，如形态学变化、DNA损伤和断裂、线粒体功能障碍、细胞色素c释放以及caspase-3和-9原表达增加。DAS还促进AIF和Endo G从HeLa细胞线粒体中的释放。总之，DAS诱导G 0/G1期细胞周期停滞和凋亡的HeLa细胞通过半胱天冬酶和线粒体和p53通路提供进一步了解DAS的作用在宫颈癌的分子机制。因此，本研究揭示DAS在体外显著抑制人宫颈癌HeLa细胞的生长并诱导其凋亡。
Diallyl sulfide (DAS), one of the main active constituents of garlic, causes growth inhibition of cancer cells in vitro and promotes immune responses in vivo in experimental settings. However, its effects on the induction of cell cycle and apoptosis in human cervical cancer cells are still unclear. The aims of this study were to explore the anti-cancer effects of DAS in He La human cervical cancer cells and to investigate the underlying mechanisms in vitro. Cytotoxicity and apoptosis in He La human cervical cancer cells were examined by the morphological changes, viability assay, 4',6-Diamidino-2-phenylindole dihydrochloride (DAPI) staining, comet assay, Western blotting and confocal microscopy examination. The results showed that DAS treatment for 24-72 h resulted in a marked decrease in cell viability time- and dose-dependently. Flow cytometric analysis showed that a 48-h treatment of 75 mu M DAS induced G0/G1 cell cycle arrest and sub-G1 phase (apoptosis) in He La cells. Typical apoptotic nucleus alterations were observed by fluorescence microscopy in He La cells after exposure to DAS using DAPI staining. Cells treated with different concentrations of DAS also showed changes typical of apoptosis such as morphological changes, DNA damage and fragmentation, dysfunction of mitochondria, cytochrome c release and increased expression of pro-caspase-3 and -9. DAS also promoted the release of AIF and Endo G from mitochondria in HeLa cells. In conclusion, DAS induced G0/G1 cell cycle arrest and apoptosis in HeLa cells through caspase- and mitochondria and p53 pathways providing further understanding of the molecular mechanisms of DAS action in cervical cancer. This study, therefore, revealed that DAS significantly inhibits the growth and induces apoptosis of human cervical cancer HeLa cells in vitro.