Chinese herbal medicine Fuzheng Kang-Ai decoction sensitized the effect of gefitinib on inhibition of human lung cancer cells through inactivating PI3-K/Akt -mediated suppressing MUC1 expression
Chinese herbal medicine Fuzheng Kang-Ai decoction sensitized the effect of gefitinib on inhibition of human lung cancer cells through inactivating PI3-K/Akt -mediated suppressing MUC1 expression
复制标题
中草药扶正抗艾汤通过灭活PI3-K/Akt介导的MUC1表达来增敏吉非替尼抑制人肺癌细胞的作用
DOI：
10.1016/j.jep.2016.10.077
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发表时间：
2016-12-24
影响因子：
5.4
通讯作者：
Hann, Swei Sunny
中科院分区：
文献类型：
--
作者：
Li, Longmei;Wang, SuMei;Hann, Swei Sunny
民族药理学相关性：中药扶正抗癌汤作为辅助治疗肺癌已有几十年的历史。然而，EGFR-TKI吉非替尼的增敏作用的分子机制尚未完全阐明。材料与方法：MTT法检测细胞活力。流式细胞仪检测细胞周期分布。Western blot检测Akt、p65、p50和MUC 1的磷酸化和蛋白水平。通过qRT-PCR测量MUC 1的mRNA水平。瞬时转染实验用于Akt、p65和MUC 1的过表达。结果：FZKA对肺癌细胞的生长有抑制作用，且FZKA与吉非替尼联合收割机联合处理对肺癌细胞的生长抑制作用更明显。FZKA可使细胞阻滞于G 0/G1期。从机制上讲，我们发现FZKA抑制Akt的磷酸化，p65和MUC 1的蛋白表达，并且在FZKA和吉非替尼组合中观察到更多的反应。过表达Akt可克服FZKA对p65蛋白表达的抑制作用，而外源性表达p65可对抗FZKA对MUC 1蛋白表达的抑制作用。相反，虽然过表达MUC 1对p65表达没有影响，但它反馈增加Akt的磷酸化，更重要的是，逆转了FZKA对细胞生长的抑制作用。FZKA和吉非替尼联合应用对NSCLC细胞生长、Akt磷酸化、p65和MUC 1蛋白表达具有协同作用。结论：FZKA水煎剂通过抑制Akt介导的p65蛋白表达，进而降低MUC 1蛋白表达，抑制NSCLC细胞生长。更重要的是，FZKA煎剂与吉非替尼合用具有协同作用，抑制作用更强。MUC 1对Akt信号通路的正反馈调节环进一步增加了MUC 1在介导FZKA水煎剂在此过程中的整体反应中的重要作用。体外和体内研究提供了一个额外的和新的机制，FZKA煎剂增强吉非替尼在吉非替尼耐药NSCLC细胞中的生长抑制。
Ethnopharmacological relevance: Chinese herbal medicine (CHM) Fuzheng Kang-Ai (FZKA for short) decoction has been used as adjuvant treatment strategies in lung cancer patients for decades. However, the molecular mechanism underlying the therapeutic potential especially in sensitizing the effect of EGFR-TKI gefitinib has not been well elucidated.Materials and methods: Cell viability was detected by MTT assay. Cell cycle distribution was detected by flow cytometry. Western blot were used to examine phosphorylation and protein levels of Akt, p65, p50 and MUC1. The mRNA level of MUC1 was measured by qRT-PCR. Transient transfection experiments were used to overexpression of Akt, p65 and MUC1. Tumor xenograft and bioluminescent imaging experiments were carried out to confirm the in vitro findings.Results: Cell viability was inhibited by FZKA treatment and showed more significant when treated with FZKA and gefitinib in combine in lung cancer cells. FZKA induced the cell arrest at G0/G1 phase. Mechanistically, we showed that the phosphorylation of Akt, protein expressions of p65 and MUC1 were suppressed by FZKA and even more responses were observed in the FZKA and gefitinib combining. Overexpressed Akt overcame the effect of FZKA on p65 protein, and exogenously expressed p65 resisted the inhibitory effect of MUC1 protein expression by FZKA. On the contrary, while overexpressed MUC1 had no effect on p65 expression, it feedback increased phosphorylation of Akt, and more importantly, reversed the cell growth inhibition affected by FZKA. In line with the above, our results confirmed the synergistic effects of FZKA and gefitinib combination on tumor growth, the phosphorylation of Akt, and protein expression of p65 and MUC1 in vivo.Conclusion: This study shows that FZKA decoction inhibits the growth of NSCLC cells through Akt-mediated inhibition of p65, followed by reducing the expression of MUC1. More importantly, there is a synergistic effect of FZKA decoction and gefitinib combination with greater suppression. The positive feedback regulatory loop of MUC1 to Akt signaling pathway further added the important role of MUC1 in mediating the overall responses of FZKA decoction in this process. The in vitro and in vivo study provides an additional and a novel mechanism by which the FZKA decoction enhances the growth inhibition of gefitinib in gefitinib-resistant NSCLC cells.