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中国临床药理学与治疗学 ›› 2004, Vol. 9 ›› Issue (9): 983-987.

• 研究原著 • 上一篇    下一篇

p42/44 MAPK 激酶抑制剂增强二烯丙基二硫化物诱导CNE2 细胞凋亡

文军, 张懿玮, 聂亚莉, 徐明   

  1. 中南大学分子药物与治疗研究所, 长沙410078, 湖南
  • 收稿日期:2003-06-12 修回日期:2004-05-08 出版日期:2004-09-26 发布日期:2020-11-23
  • 通讯作者: 徐明, 男, 博士, 教授, 硕士生导师, 主要从事分子医学研究。Tel:0731-2685181 E-mail:mjs xu @yahoo. com
  • 作者简介:文军, 男, 硕士, 讲师。Tel:0731-4364333 E-mail:jun wen719 @yahoo.com. cn
  • 基金资助:
    中南大学归国留学人员专项基金(№181176059)

Inhibition of p42/44 MAPK kinase enhances diallyl disulfide induced apoptosis in human CNE2 cells

WEN Jun, Zhang Yi Wei, Nie Ya-Li, XU Ming   

  1. Research Institute for Molecular Pharmacology and Therapeutics, Centralsouth University, Changsha 410078, Hunan, China
  • Received:2003-06-12 Revised:2004-05-08 Online:2004-09-26 Published:2020-11-23

摘要: 目的: 探讨二烯丙基二硫化物(DADS) 诱导CNE2 细胞凋亡及p42/44 MAPK 信号转导通路对此过程的作用。方法: DADS 处理CNE2 细胞24 h 后, 荧光显微镜下观察形态学变化及凋亡细胞计数, 四甲基偶氮唑盐微量酶反应比色法测定细胞活性, 流式细胞仪检测凋亡细胞, 蛋白质印迹法检测磷酸化p42/44MAPK 表达。结果: DADS(50 ~ 150 μmol·L-1) 作用24 h 后, 诱导CNE2 细胞产生典型的凋亡细胞形态学变化(核浓染核碎裂), 流式细胞仪结果显示, 随着DADS 给药浓度增加, 细胞凋亡率呈浓度依赖性逐渐升高, 细胞周期中各期细胞所占百分率的变化无规律, DADS(50 ~ 150 μmol·L-1 ) 浓度依赖性刺激磷酸化p42/44 MAPK 的表达, p42/44 MAPK 抑制剂U0126 显著增强DADS 致凋亡作用。结论: DADS 诱导CNE2 细胞凋亡时激活磷酸化p42/44 MAPK 表达, 磷酸化p42/44 MAPK 抑制剂能增强DADS 诱导CNE2 细胞凋亡效应。

关键词: 细胞凋亡, 磷酸化p42/44 MAPK, 二烯丙基二硫化物, 蛋白质印迹, CNE2 细胞

Abstract: AIM: To investigate the effects of p42/44 MAPK signaling on diallyl disulfide-induced apoptosis in CNE2 cells.METHODS: Morphological changes and quantification of apoptotic cells were determined under fluorescence microscope after a 24 h treatment of CNE2 by Diallyl disulfide (DADS ). Cell viability was determined withMTT method. Apoptosis detection which was taken to the cells on flowcytometry. The expression of phosphorp42 44 MAPK was measured by Western blotting.RESULTS: After treatment with DADS at 50-150 μmol·L-1 for 24 h, DADS elicited typical apoptotic morphologic changes (chromatic condensation and nucleus fragmentation). The amount of apoptosis cells increased in a concentration-dependent manner but cell-cycle arrest was not. At the concentration between 50 and 150 μmol·L-1, incubation of CNE2 with DADS for 24 h also induced phosphor-p42/44 MAPK expression in a concentration-dependent manner. Interestingly, DADS induced apoptosis was markedly increased by preincubation with U0126, a specific p42/44MAPK inhibitor, and increased the expression of phosphor-p42/44 MAPK in a concentration-dependent manner.CONCLUSION: DADS activates p42/44MAPK, and phosphor-p42/44MAPK signaling appears to play protective roles in DADS-induced apoptosis in CNE2.

Key words: apoptosis, phospho-p42/44 MAPK, diallyl disulfide, Western blotting, CNE2

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