田颖颖;杨爱琳;陈孝男;任浩淼;刘亚鑫;邱海玲;唐雷梦媛;黄惠铭;屠鹏飞;胡仲冬;
TIAN Ying-ying;YANG Ai-lin;CHEN Xiao-nan;REN Hao-miao;LIU Ya-xin;QIU Hai-ling;TANG Lei-meng-yuan;HUANG Hui-ming;TU Peng-fei;HU Zhong-dong;Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine;

• 1:北京中医药大学中药学院中药现代研究中心

摘要(Abstract)：

探讨槐耳清膏对人非小细胞肺癌NCI-H1299细胞生长和转移的抑制作用及其潜在的作用机制。采用细胞增殖毒性试验(MTT法)检测槐耳清膏对NCI-H1299细胞增殖的影响。采用流式细胞术检测槐耳清膏给药后对NCI-H1299细胞凋亡率、细胞周期以及细胞内活性氧(ROS)水平的影响。采用细胞划痕试验研究槐耳清膏给药后对NCI-H1299细胞迁移能力的影响。此外,借助免疫印迹法检测槐耳清膏给药后与凋亡和上皮-间质转化(epithelial-mesenchymal transition,EMT)过程相关的蛋白以及MAPK信号通路中的关键蛋白表达水平的变化。研究结果显示,槐耳清膏能够抑制人非小细胞肺癌NCI-H1299细胞的增殖,而且诱导细胞周期阻滞在S期。槐耳清膏能够通过下调抗凋亡蛋白Bcl-2的表达促进NCI-H1299细胞凋亡的发生。此外,槐耳清膏给药能够升高NCI-H1299细胞内ROS的含量从而诱导细胞铁死亡的发生。EMT在癌症转移中起着重要作用。槐耳清膏给药后能够降低NCI-H1299细胞的迁移能力,并且能够下调间质标志物TCF8/ZEB1,N-cadherin,β-catenin的蛋白水平进而抑制NCI-H1299细胞EMT进程。该研究还揭示在人非小细胞肺癌NCI-H1299细胞中槐耳清膏能够抑制MAPK信号通路,这可能是槐耳清膏抑制NCI-H1299细胞生长和转移的机制之一。该项工作为槐耳抗肺癌作用研究提供了新的内容,也为槐耳临床治疗肺癌提供了新的理论依据,对后续深入研究槐耳抗肿瘤作用机制具有一定的参考意义。
This study aims to investigate the effect of Huaier aqueous extract on the growth and metastasis of human non-small cell lung cancer NCI-H1299 cells and its underlying mechanisms. MTT assay was used to detect the effect of Huaier aqueous extract on the proliferation of NCI-H1299 cells. Flow cytometry was used to examine the effect of Huaier aqueous extract on the apoptosis, cell cycle, and ROS level of NCI-H1299 cells. Wound healing assay was used to evaluate the effect of Huaier aqueous extract on the migration ability of NCI-H1299 cells. Western blot was used to detect the levels of proteins involving apoptosis, epithelial-mesenchymal transition(EMT), and MAPK signaling pathway in NCI-H1299 cells exposed to Huaier aqueous extract. The results showed that Huaier aqueous extract inhibited the proliferation of NCI-H1299 cells, and induced cell-cycle arrest at the phase S. Huaier aqueous extract promoted the apoptosis of NCI-H1299 cells by down-regulating the expression of anti-apoptotic protein Bcl-2. Moreover, Huaier aqueous extract increased ROS level and induced ferroptosis in NCI-H1299 cells. EMT played a critical role in cancer metastasis. Huaier aqueous extract reduced the migration ability of NCI-H1299 cells by inhibiting EMT of NCI-H1299 cells. In addition, this study revealed that Huaier aqueous extract inhibited MAPK signaling pathway in human non-small cell lung cancer NCI-H1299 cells, which may be one of Huaier′s mechanisms in inhibiting growth and metastasis of NCI-H1299 cells. This study provides a new theoretical basis for the clinical treatment of lung cancer with Huaier, and important reference significance for further studies on the anti-tumor mechanisms of Huaier.

关键词(KeyWords)： 槐耳清膏;人非小细胞肺癌细胞;增殖;转移;凋亡;铁死亡
Huaier aqueous extract;human non-small cell lung cancer cells;proliferation;metastasis;apoptosis;ferroptosis

Abstract:

Keywords:

基金项目(Foundation): 中华中医药学会青年人才托举工程项目(CACM-2018-(QNRC2-B05));; 北京市科技新星计划项目(Z191100001119083);; 北京中医药大学大学生创新创业训练项目(201910026087)

作者(Authors): 田颖颖;杨爱琳;陈孝男;任浩淼;刘亚鑫;邱海玲;唐雷梦媛;黄惠铭;屠鹏飞;胡仲冬;
TIAN Ying-ying;YANG Ai-lin;CHEN Xiao-nan;REN Hao-miao;LIU Ya-xin;QIU Hai-ling;TANG Lei-meng-yuan;HUANG Hui-ming;TU Peng-fei;HU Zhong-dong;Modern Research Center for Traditional Chinese Medicine, School of Chinese Materia Medica, Beijing University of Chinese Medicine;

DOI: 10.19540/j.cnki.cjcmm.20200310.401

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