杠柳乙酸乙酯提取物通过激活 Hippo 信号通路和抑制 YAP 核转位抑制非小细胞肺癌。
The Ethyl acetate extract from Celastrus orbiculatus suppresses non-small-cell lung cancer by activating Hippo signaling and inhibiting YAP nuclear translocation.
机构信息
Department of Respiratory Medicine, The Affiliated Hospital of Yangzhou University, Yangzhou University, Yangzhou, Jiangsu, PR China.
Yangzhou Hospital of Traditional Chinese Medicine, Yangzhou, 225001, PR China.
出版信息
Phytomedicine. 2023 Jun;114:154761. doi: 10.1016/j.phymed.2023.154761. Epub 2023 Mar 12.
BACKGROUND
Celastrus orbiculatus Thunb. is a medicinal plant that has been widely used for thousands of years in China, and the ethyl acetate extract (Celastrus orbiculatus Thunb. Extract, COE) from its stem was reported to exert antitumor and anti-inflammatory effects in various preclinical studies. However, the anti-non-small-cell lung cancer activity of COE and its potential mechanism are not yet fully understood.
PURPOSE
To investigate the antitumor effects of COE on non-small-cell lung cancer (NSCLC) cells and explore its molecular mechanism from the perspective of Hippo signaling, YAP nuclear translocation, and reactive oxygen species (ROS) generation.
METHODS
The effects of COE on proliferation, cell cycle arrest, apoptosis, stemness, and senescence in NSCLC cell lines were determined by CCK-8, clone formation, flow cytometry, and β-galactosidase staining assays. The effects of COE on Hippo signaling were investigated by Western blotting. The intracellular expression and distribution of YAP were analyzed by immunofluorescence assay. DCFH-DA probe combined with flow cytometry was used to detect intracellular total ROS levels in NSCLC cells after COE treatment. Xenograft tumor model was established, and the animal living image system was employed to analyze the effects of COE on the Hippo-YAP signaling in vivo.
RESULT
COE significantly inhibited NSCLC activity in vitro and in vivo, mainly by proliferation inhibition, cycle arrest, apoptosis promotion, senescence promotion, and stemness downregulation. COE strongly activated Hippo signaling and inhibited YAP expression and nuclear retention. Activation of Hippo signaling induced by COE was associated with ROS-mediated phosphorylation of MOB1.
CONCLUSION
This study demonstrated that COE inhibited NSCLC through activating Hippo signaling and suppressing YAP nuclear translocation, in which ROS may play a role in the phosphorylation of the MOB1 protein.
背景
南蛇藤是一种药用植物，在中国已被广泛应用数千年，其茎的乙酸乙酯提取物（南蛇藤提取物，COE）在各种临床前研究中显示出抗肿瘤和抗炎作用。然而，COE 对非小细胞肺癌（NSCLC）的抗肿瘤活性及其潜在机制尚未完全阐明。
目的
从 Hippo 信号、YAP 核易位和活性氧（ROS）生成的角度探讨 COE 对 NSCLC 细胞的抗肿瘤作用及其分子机制。
方法
通过 CCK-8、克隆形成、流式细胞术和β-半乳糖苷酶染色实验，测定 COE 对 NSCLC 细胞系增殖、细胞周期阻滞、凋亡、干性和衰老的影响。通过 Western blot 检测 COE 对 Hippo 信号的影响。免疫荧光检测 COE 对 YAP 细胞内表达和分布的影响。用 DCFH-DA 探针结合流式细胞术检测 COE 处理后 NSCLC 细胞内总 ROS 水平。建立异种移植肿瘤模型，利用动物活体成像系统分析 COE 对体内 Hippo-YAP 信号的影响。
结果
COE 显著抑制 NSCLC 的体内外活性，主要通过抑制增殖、细胞周期阻滞、促进凋亡、促进衰老和下调干性来实现。COE 强烈激活 Hippo 信号，抑制 YAP 的表达和核保留。COE 激活 Hippo 信号与 ROS 介导的 MOB1 磷酸化有关。
结论
本研究表明，COE 通过激活 Hippo 信号并抑制 YAP 核易位来抑制 NSCLC，其中 ROS 可能在 MOB1 蛋白的磷酸化中发挥作用。
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