A Novel Benzofuran Derivative Moracin N Induces Autophagy and Apoptosis Through ROS Generation in Lung Cancer
A Novel Benzofuran Derivative Moracin N Induces Autophagy and Apoptosis Through ROS Generation in Lung Cancer
复制标题
新型苯并呋喃衍生物 Moracin N 通过 ROS 生成在肺癌中诱导自噬和细胞凋亡
DOI：
10.3389/fphar.2020.00391
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发表时间：
2020-05-12
影响因子：
5.6
通讯作者：
Tian, Jingkui
中科院分区：
文献类型：
--
作者：
Gao, Chengcheng;Sun, Xin;Tian, Jingkui
前言桑叶是一种传统中药，广泛应用于肺部疾病。莫拉克因N（MAN）是从桑叶中提取的一种次生代谢产物，是一种有效的抗癌药物。但其分子机制仍不清楚。目的研究MAN对人肺癌的作用，并探讨其分子机制。方法MTT法检测细胞活力。Annexin V-FITC/PI染色检测细胞凋亡。共聚焦显微镜观察自噬体和自溶体的形成。进行流式细胞术以定量细胞死亡。Western blotting检测相关信号通路。结果本研究首次证实MAN抑制人非小细胞肺癌（NSCLC）细胞增殖并诱导细胞凋亡。我们发现MAN处理使A549和PC 9细胞的线粒体功能失调并导致线粒体凋亡。同时，MAN通过增加自噬体的形成、自噬体与溶酶体的融合以及溶酶体的功能来增强自噬通量。MAN对自噬的经典调控途径mTOR信号通路有时间和剂量依赖性的抑制作用，从而诱导自噬。有趣的是，通过CQ或Atg 5敲低的自噬抑制减弱了MAN引起的细胞凋亡，表明自噬起到细胞死亡的作用。此外，自噬介导的细胞死亡的MAN可以阻断活性氧（ROS）清除剂NAC，表明ROS积累是诱导因素的凋亡和自噬。总之，我们揭示了MAN通过细胞凋亡和自噬抗肺癌的分子机制，提示MAN可能是一种新型的治疗NSCLC的药物。
Introduction The leaves of Morus alba L is a traditional Chinese medicine widely applied in lung diseases. Moracin N (MAN), a secondary metabolite extracted form the leaves of Morus alba L, is a potent anticancer agent. But its molecular mechanism remains unveiled. Objective In this study, we aimed to examine the effect of MAN on human lung cancer and reveal the underlying molecular mechanism. Methods MTT assay was conducted to measure cell viability. Annexin V-FITC/PI staining was used to detect cell apoptosis. Confocal microscope was performed to determine the formation of autophagosomes and autolysosomes. Flow cytometry was performed to quantify cell death. Western blotting was used to determine the related-signaling pathway. Results In the present study, we demonstrated for the first time that MAN inhibitd cell proliferation and induced cell apoptosis in human non-small-cell lung carcinoma (NSCLC) cells. We found that MAN treatment dysregulated mitochondrial function and led to mitochondrial apoptosis in A549 and PC9 cells. Meanwhile, MAN enhanced autophagy flux by the increase of autophagosome formation, the fusion of autophagsomes and lysosomes and lysosomal function. Moreover, mTOR signaling pathway, a classical pathway regualting autophagy, was inhibited by MAN in a time- and dose-dependent mannner, resulting in autophagy induction. Interestingly, autophagy inhibition by CQ or Atg5 knockdown attenuated cell apoptosis by MAN, indicating that autophagy serves as cell death. Furthermore, autophagy-mediated cell death by MAN can be blocked by reactive oxygen species (ROS) scavenger NAC, indicating that ROS accumulation is the inducing factor of apoptosis and autophagy. In summary, we revealed the molecular mechanism of MAN against lung cancer through apoptosis and autophagy, suggesting that MAN might be a novel therapeutic agent for NSCLC treatment.