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去甲斑蝥素对人肺腺癌A549细胞的抑制作用

崔宝弟,王  敏#,孙震晓*   

  1. 北京中医药大学中药学院生物制药系,北京  100102
  • 收稿日期:2014-06-22 修回日期:2014-09-23 出版日期:2015-01-31 发布日期:2015-01-31
  • 通讯作者: 孙震晓,Tel:010-84738646; E-mail:sunzxcn@hotmail.com
  • 作者简介:崔宝弟,E-mail:cbdbaodi@sina.com。#该作者对本文的贡献与第一作者等同
  • 基金资助:

    北京中医药大学自主课题项目(2014JYBZZ-XS097);北京中医药大学自主课题项目(2013JYBzz-XS088)

The inhibition of norcantharidin on human lung adenocarcinoma A549 cells

CUI Baodi,WANG Min#,SUN Zhenxiao*   

  1. Department of Biopharmaceutical, School of Chinese Pharmacology, Beijing University of Chinese Medicine, Beijing 100102, China
  • Received:2014-06-22 Revised:2014-09-23 Online:2015-01-31 Published:2015-01-31

摘要:

目的: 研究去甲斑蝥素(NCTD)对人肺腺癌A549细胞的细胞毒作用,并初步探讨其作用机制。方法:采用四甲基噻唑蓝(MTT)试验检测0~240 μmol/L NCTD在0~72 h内对A549细胞活力的影响;NCTD处理A549细胞24 h后换正常培养基,用MTT方法检测细胞恢复与增殖的情况;采用倒置显微镜观察A549细胞经40、50和60 μmol/L NCTD处理0~72 h后细胞形态学的改变;通过流式细胞术检测40~60 μmol/L NCTD对细胞周期和凋亡的影响。结果:30~240 μmol/L NCTD对A549细胞有明显的生长抑制作用(P<0.05);在30~120 μmol/L浓度范围内,NCTD对A549细胞的生长抑制作用在NCTD撤除24 h后逐渐消失,30~60 μmol/L NCTD作用A549细胞24 h后,A549细胞的活力在恢复培养的第5天完全恢复;40、50和60 μmol/L NCTD作用A549细胞0~72 h,细胞表现为明显的G2~M期阻滞,与对照组相比,NCTD诱导A549细胞凋亡作用不明显(P>0.05)。结论:40~60 μmol/L NCTD主要通过诱导A549细胞G2~M期阻滞而抑制细胞生长。

关键词: 去甲斑蝥素, 肺癌, A549细胞, 细胞周期, 细胞凋亡

Abstract:

OBJECTIVE: To study the effects of norcantharidin (NCTD) on human lung cancer cells,and investigate the mechanisms. METHODS:The growth inhibition of A549 cells treated with 0-240 μmol/L NCTD for 0-72 hours was analyzed by MTT assay. The recovery growth and proliferation of A549 cells treated with 0-120 μmol/L NCTD for 24 h was evaluated by MTT assay. The morphological changes of A549 cells treated with 40,50 and 60 μmol/L NCTD for 0-72 h were examined under inverted microscope. The apoptosis and cell cycle changes of A549 cells treated with 40-60 μmol/L NCTD were detected by flow cytometry. RESULTS:NCTD inhibited the growth of A549 cells in 30-240 μmol/L(P<0.05). Cell growth inhibition could hardly be detected 24 h after 30-240 μmol/L NCTD was removed from culture medium. The vitality of A549 cells was fully recovered in 5 d after 30-60 μmol/L NCTD was removed from culture medium. A549 cells showed G2-M phase block but no obvious apoptosis compared with the control group when treated with 40,50 and 60 μmol/L NCTD for 0-72 h. CONCLUSION: NCTD (40-60 μmol/L) inhibited human lung cancer A549 cell growth mainly by cell G2-M phase block.

Key words: norcantharidin, lung cancer, A549 cell, cell cycle, apoptosis