广西师范大学学报（自然科学版） ›› 2015, Vol. 33 ›› Issue (3): 91-97.doi: 10.16088/j.issn.1001-6600.2015.03.014

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Hsp90抑制剂木犀草苷对非小细胞肺癌的体外抑制活性及作用机理研究

梅寒冰1,2, 王威1,2, 姚雪1,2, 程联彪1,2, 黄瑾1,2   

  1. 1. 华东理工大学药学院,上海200237;
    2. 华东理工大学新药设计上海市重点实验室,上海200237
  • 收稿日期:2015-04-19 出版日期:2015-05-10 发布日期:2018-09-20
  • 通讯作者: 黄瑾(1976—),女,广西桂林人,华东理工大学教授,博士。E-mail: huangjin@ecust.edu.cn
  • 基金资助:
    国家自然科学基金资助项目(91313303)

Inhibitory Activity and Mechanism of Hsp90 Inhibitor Cynaroside Against Non-small Cell Lung Cancer in Vitro

MEI Han-bing1,2, WANG Wei1,2, YAO Xue1,2, CHENG Lian-biao1,2, HUANG Jin1,2   

  1. 1. School of Pharmacy, East China University of Science and Technology, Shanghai 200237, China;
    2. Shanghai Key Laboratory of New Drug Design, East China University of Science and Technology,Shanghai 200237, China
  • Received:2015-04-19 Online:2015-05-10 Published:2018-09-20

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摘要： 本文利用荧光偏振法(FP)构建分子伴侣Hsp90的筛选平台,筛选得到黄酮类化合物木犀草苷,并用热稳定实验验证了它与蛋白的结合作用;评价了木犀草苷对14株癌细胞的体外增殖抑制作用,发现木犀草苷对非小细胞肺癌有良好的选择性抑制活性;利用细胞克隆形成、western blot以及流式细胞检测技术进一步深入研究其可能的作用机制,发现木犀草苷能剂量依赖性地抑制非小细胞肺癌的增殖;下调Hsp90的客户蛋白Raf-1、HER2的表达;将H460细胞阻滞在G2/M期;诱导细胞凋亡,并且引起Bax的表达上调。实验结果表明,黄酮类化合物木犀草苷可以通过抑制Hsp90的活性进而降解肿瘤细胞内的客户蛋白,诱导H460细胞G2/M期阻滞,上调Bax表达诱导细胞凋亡来发挥抗肿瘤活性。

关键词: Hsp90, 木犀草苷, 非小细胞肺癌, 细胞周期阻滞, 凋亡

Abstract: A fluorescence polarization technique was established to screen Hsp90 inhibitors and was selected as candidate for further anti-proliferation study. Thermal shift assay was used to confirm the binding effect of cynaroside and Hsp90. Then it was found that cynaroside showed potent and selective antiproliferation activities against non-small lung cancer cells among 14 strains of cancer cells. The colonial assay, western blot and flow cytometry were employed to investigate its possible mechanism. The results showed that cynaroside could inhibit the proliferation of non-small lung cancer cells in a dose-dependent manner and induce the degradation of Raf-1, and HER2 expression. Moreover, cynaroside induced G2/M arrest in H460 cell and triggered apoptosis by up-regulatiing Bax expression. Finally, the results displayed that cynaroside, one of the flavonoids, formed antineoplastic activity by inhibiting Hsp90 activity to degrade its client proteins in H460 cells, inducing G2/M phase arrest and triggering apoptosis by up-regulating Bax expression.

Key words: Hsp90, cynaroside, non-small lung cancer, cell cycle arrest, apoptosis

中图分类号: 

  • R962
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