Daphnetin triggers ROS-induced cell death and induces cytoprotective autophagy by modulating the AMPK/Akt/mTOR pathway in ovarian cancer
Daphnetin triggers ROS-induced cell death and induces cytoprotective autophagy by modulating the AMPK/Akt/mTOR pathway in ovarian cancer
复制标题
瑞香素通过调节卵巢癌中的 AMPK/Akt/mTOR 通路触发 ROS 诱导的细胞死亡并诱导细胞保护性自噬
DOI：
10.1016/j.phymed.2021.153465
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发表时间：
2021-01-21
期刊：
影响因子：
7.9
通讯作者：
Zhang, Songling
中科院分区：
文献类型：
--
作者：
Fan, Xiaoye;Xie, Min;Zhang, Songling
背景：卵巢癌是世界上最常见的妇科恶性肿瘤之一。瑞香素（Daphnetin，Daph）是一种具有抗肿瘤活性的化合物，但其抗卵巢癌作用的分子机制尚不清楚。目的：本研究旨在探讨Daph对卵巢癌细胞的抗肿瘤作用及其机制。细胞生长、增殖、凋亡和ROS产生通过CCK 8测定、集落形成测定和流式细胞术测量。Western blotting检测相关信号蛋白。免疫荧光和透射电子显微镜被用来评估自噬和自噬通量的标志物。在A2780异种移植模型中观察到抗肿瘤作用。结果：Daph通过诱导卵巢癌细胞ROS依赖性凋亡发挥抗肿瘤作用，并可被NAC逆转。AMPK/Akt/mTOR信号通路参与Daphs介导的细胞保护性自噬，当Daphs介导的AMPK表达水平和自噬被阻断时，细胞增殖受到强烈抑制，细胞凋亡受到诱导。此外，在A2780异种移植模型中，Daph和自噬抑制剂的联合治疗显示出明显的协同作用，抑制细胞活力和促进凋亡，没有任何副作用。结论：我们的研究结果表明，Daph触发ROS诱导的细胞凋亡，并通过调节AMPK/Akt/mTOR通路诱导细胞保护性自噬。此外，Daph和自噬抑制剂的组合可能是卵巢癌的潜在治疗策略。
Background: Ovarian cancer is one of the most common gynecological malignancies in the world. Daphnetin (Daph) was previously reported to possess antitumor potential, but its potential and molecular mechanisms in ovarian cancer remain poorly understood.Purpose: In the current study, we aimed to explore the antitumor effect and detailed mechanisms of Daph in ovarian cancer cells.Methods: The cytotoxic effect of Daph on ovarian cells was determined in vitro and in vivo. Cell growth, proliferation, apoptosis and ROS generation were measured by CCK8 assays, colony formation assays and flow cytometry. Western blotting was used to evaluate the related signal proteins. Immunofluorescence and transmission electron microscopy were used to evaluate markers of autophagy and autophagic flux. The antitumor effects were observed in the A2780 xenograft model. Moreover, Daph-induced autophagy was observed by enhanced LC3-II accumulation and endogenous LC3 puncta, and an autophagy inhibitor further enhanced the antitumor efficacy of Daph, which indicated that the cytoprotective role of autophagy in ovarian cancer.Results: We found that Daph exhibited antitumor effects by inducing ROS-dependent apoptosis in ovarian cancer, which could be reversed by N-acetyl cysteine (NAC). The AMPK/Akt/mTOR pathway was involved in Daph-mediated cytoprotective autophagy, and when Daph-mediated the expression level of AMPK and autophagy were blocked, there was robust inhibition of cell proliferation and induction of apoptosis. In addition, in the A2780 xenograft model, combined treatment with Daph and an autophagy inhibitor showed obvious synergetic effects on the inhibition of cell viability and promotion of apoptosis, without any side effects.Conclusion: Our results suggest that Daph triggers ROS-induced cell apoptosis and induces cytoprotective autophagy by modulating the AMPK/Akt/mTOR pathway. Moreover, the combination of Daph and autophagy inhibitor may be a potential therapeutic strategy for ovarian cancer.