Traditional Chinese medicine Ze-Qi-Tang formula inhibit growth of non-small-cell lung cancer cells through the p53 pathway
Traditional Chinese medicine Ze-Qi-Tang formula inhibit growth of non-small-cell lung cancer cells through the p53 pathway
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中药泽气汤方通过p53通路抑制非小细胞肺癌细胞生长
DOI：
10.1016/j.jep.2019.01.007
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发表时间：
2019-04-24
影响因子：
5.4
通讯作者：
Zou, Chunpu
中科院分区：
文献类型：
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作者：
Xu, Zihang;Zhang, Fei;Zou, Chunpu
民族药理学相关性：泽芪汤（ZQT）是一种经典的中药方剂，用于治疗咳嗽、哮喘、胸腔积液和肺癌等多种呼吸系统疾病已有数千年的历史。研究目的：ZQT中药方剂的累积文献显示，其几种组成成分是不同癌细胞凋亡的强效诱导剂。然而，ZQT对非小细胞肺癌（NSCLC）的活性以前尚未研究过。材料与方法：采用CCK-8法和集落形成实验检测细胞生长情况，并与对照组比较，探讨ZQT对NSCLC细胞生长的影响。采用Annexin V/碘化丙啶、Hoechst 33342或TUNEL染色法，通过流式细胞术分析确定细胞凋亡诱导或细胞周期阻滞。在一些测定中，在用ZQT处理之前，用匹非亭-α阻断NSCLC（A549和H460）细胞的p53活性。Western blot检测细胞周期和凋亡相关蛋白的表达水平。ZQT（100 mg/kg、200 mg/kg、400 mg/kg）对皮下接种A549细胞的裸鼠体内抗肿瘤活性进行了检测。每周测量小鼠的体重和肿瘤体积。生存印记被记录下来。TUNEL法检测小鼠组织细胞凋亡。结果：ZQT对NSCLC（A549和H460）细胞的增殖抑制呈剂量和时间依赖性，并沿着诱导细胞周期阻滞于G 0/G1期。细胞周期阻滞的阻滞和细胞增殖的抑制与p53沿着上调以及细胞周期蛋白B1和Cdk 2的下调相关，表明ZQT诱导线粒体相关的凋亡。ZQT诱导的细胞凋亡和G 0/G1期阻滞的逆转观察与pifithrin-a预处理。还发现ZQT抑制小鼠异种移植模型中肿瘤生长的进展并延长存活。此外，没有肝或肾或心脏毒性与ZQT治疗检测在mice.Conclusion：这些研究结果表明，ZQT配方抑制非小细胞肺癌细胞的生长，是一个潜在的代理人的补充和替代治疗肺癌。
Ethnopharmacological relevance: Ze-Qi-Tang (ZQT), a classic Chinese herbal formula, has been for over thousand years used for the treatment of several respiratory ailments like cough, asthma, hydrothorax and lung cancer.Aim of study: Cumulative literature on ZQT herbal formula reveals that its several constituent components are potent inducer of apoptosis in different cancer cells. However, the activity of ZQT against non-small-cell-lung cancer (NSCLC) has not been previously examined. The aim of the study is to investigate the molecular mechanism of ZQT on NSCLC cells.Materials and methods: Cell growth were determined by CCK-8 and colony formation assay. Induction of cellular apoptosis or arrest of cell cycle were determined by flow cytometric analysis using annexin V/ propidium iodide, Hoechst 33342 or TUNEL staining method. In some assay p53 activity of NSCLC ( A549 and H460) cells were blocked with pifithrin-a, prior to treatment with ZQT. The level of expression of cell cycle and apoptosis related marker proteins were estimated by western blot. The anticancer activity of ZQT in vivo were monitored in nude mice that were induced with tumor by subcutaneous inoculation of A549 cells and then treated by ZQT(100 mg/kg,200 mg/kg,400 mg/kg) gavaging for 30 days. Mice' body weight and tumor volume were measured weekly. The survival carve was recorded. Apoptosis from mice' tissue was observed by TUNEL assay. Pathological histology of liver, kidney and heart were detected by H&E staining, and its functions were tested by ELISA.Results: Dose- and time-dependent inhibition of proliferation of NSCLC ( A549 and H460) cells by ZQT therapy along with induction of cell cycle arrest at G0/G1 phase were observed. The arrest of cell cycle arrest and inhibition of cellular proliferation were associated with up regulation of p53 along with down regulation of Cyclin B1 and Cdk2 indicating a mitochondrial related induction of apoptosis with ZQT. A reversal of ZQT-induced apoptosis and G0/G1 arrest was observed with pifithrin-a pretreatment. ZQT was also found to suppress the progression of tumor growth in mouse xenograft models and prolong survival. In addition, no hepato- or nephro- or cardio-toxicity with ZQT treatment were detected in mice.Conclusion: These findings suggest that the ZQT formula inhibits the growth of NSCLC cells and is a potential agent of complementary and alternative treatment for lung cancer.