实用肿瘤学杂志 ›› 2013, Vol. 27 ›› Issue (4): 344-349.doi: 10.3969/j.issn.1002-3070.2013.04.011

• 论著 • 上一篇    下一篇

丹皮酚诱导人喉癌Hep-2细胞凋亡的体外研究

隋颖, 康健, 王晓光   

  1. 中国人民解放军海军总医院耳鼻喉科(北京 100048)
  • 收稿日期:2013-02-15 出版日期:2013-08-28 发布日期:2013-08-13
  • 通讯作者: 隋颖, E-mail:zishuijingqaz@163.com
  • 作者简介:隋颖, 女, (1981-), 硕士, 住院医师, 从事头颈部肿瘤方面研究

Experimental study on apoptosis of laryngeal carcinoma Hep-2 induced by Paeonol

SUI Ying, KANG Jian, WANG Xiaoguang   

  1. PLA Center of Otolaryngology, Navy General Hospital, Beijing 100048, China
  • Received:2013-02-15 Online:2013-08-28 Published:2013-08-13

摘要: 目的研究丹皮酚对人喉癌Hep-2细胞的生长抑制作用及凋亡机制。方法 人喉癌Hep-2细胞株进行培养至对数生长期, 施加不同浓度丹皮酚(31.25mg/L、62.5mg/L、125mg/L、250mg/L)并继续培养至不同时间段, 应用四唑盐比色法(MTT)检测细胞增殖抑制率;流式细胞仪检测细胞凋亡率;荧光显微镜观察细胞凋亡形态学;Western blot检测caspase-3的表达。结果 MTT结果显示, Hep-2细胞随着丹皮酚浓度增加其体外生长受到明显的抑制。应用荧光显微镜观察发现实验组凋亡细胞数目增加。流式细胞仪检测结果显示不同浓度丹皮酚作用48h后, 细胞凋亡率不断增加, 呈剂量依赖性, 细胞周期呈现S期阻滞。Hep-2细胞随丹皮酚浓度增加和时间的延长, caspase-3蛋白的表达明显增加, 提示细胞凋亡增加。结论 丹皮酚可抑制人喉癌Hep-2细胞的体外生长, 随着丹皮酚剂量的增加及作用时间的延长, 细胞凋亡率不断增加;丹皮酚可上调caspase-3的表达, 从而诱导人喉癌Hep-2细胞凋亡。

Abstract: Objective Inhibition and apoptosis mechanism of Paeonol on human laryngeal cancer Hep-2 cell growth.Methods Cultivation to the logarithmic growth phase cells from human laryngeal carcinoma cell line Hep-2, applying different concentration of paeonol(31.25mg/L, 62.5mg/L, 125mg/L, 250mg/L)and continue to develop to different time, application of four tetrazolium colorimetric assay(MTT)to detect the inhibitory rate of cell proliferation;flow cytometry propidium iodide staining were used to detect apoptosis rate cell apoptosis was detected by fluorescence microscopy;morphology of acridine orange staining to detect the expression of Caspase-3;Western blotting technique.Results MTT Results show:Hep-2 cells with paeonol concentration increases the in vitro growth was inhibited.Flow cytometry showed different concentrations of paeonol 48h, cell apoptosis rate increased, in a dose dependent manner, the cell cycle of S phase arrest.Expression of apoptosis related gene caspase-3:Hep-2 cells with the increase of paeonol concentration and the extension of time, the expression of Caspase-3 gene increased significantly, suggesting increased apoptosis.Conclusion Paeonol could inhibit the growth of human laryngeal carcinoma Hep-2 cells in vitro, and the effect increased with the dosage of the Danpi time prolonged.apoptosis rate increased;paeonol can up-regulate the expression of Caspase-3, thereby inducing apoptosis of human laryngeal carcinoma Hep-2 cells.

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