通过抑制转移和诱导活性氧介导的凋亡和自噬细胞死亡，蛹虫草在体外和体内对人神经胶质瘤细胞的抗癌活性。
The in vitro and in vivo anticancer activities of Antrodia salmonea through inhibition of metastasis and induction of ROS-mediated apoptotic and autophagic cell death in human glioblastoma cells.
机构信息
Institute of Nutrition, College of Health Care, China Medical University, Taichung 40402, Taiwan, ROC.
Department of Cosmeceutics, College of Pharmacy, China Medical University, Taichung 40402, Taiwan, ROC; Department of Health and Nutrition Biotechnology, Asia University, Taichung 41354, Taiwan, ROC; Chinese Medicine Research Center, China Medical University, Taichung 40402, Taiwan, ROC; Research Center of Chinese Herbal Medicine, China Medical University, Taichung 40402, Taiwan, ROC.
出版信息
Biomed Pharmacother. 2023 Feb;158:114178. doi: 10.1016/j.biopha.2022.114178. Epub 2023 Jan 4.
BACKGROUND
Antrodia salmonea (AS) exhibits anticancer activities against various cancers.
OBJECTIVE
This study investigated the anticancer activities of AS on human glioblastoma (GBM8401 and U87MG) cells both in vitro and in vivo and explained the underlying molecular mechanism.
METHODS
MTT, colony formation, migration/invasion assay, immunoblotting, immunofluorescence, TUNEL, Annexin V/PI staining, AO staining, GFP-LC3 transfection, TEM, qPCR, siLC3, DCFH2-DA assay, and xenografted-nude mice were used to assess the potential of AS therapy.
RESULTS
AS treatment retarded growth and suppressed colony formation in glioblastoma cells. AS attenuates EMT by suppressing invasion and migration, increasing E-cadherin expression, decreasing Twist, Snail, and N-cadherin expression, and inhibiting Wnt/β-catenin pathways in GBM8401 and U87MG cells. Furthermore, AS induced apoptosis by activating caspase-3, cleaving PARP, and dysregulating Bax and Bcl-2 in both cell lines. TUNEL assay and Annexin V/PI staining indicated AS-mediated late apoptosis. Interestingly, AS induced autophagic cell death by LC3-II accumulation, AVO formation, autophagosome GFP-LC3 puncta, p62/SQSTM1 expression, and ATG4B inhibition in GBM8401 and U87MG cells. TEM data revealed that AS favored autophagosome and autolysosome formation. The autophagy inhibitors 3-MA/CQ and LC3 knockdown suppressed AS-induced apoptosis in glioblastoma cells, indicating that the inhibition of autophagy decreased AS-induced apoptosis. Notably, the antioxidant N-acetylcysteine (NAC) inhibited AS-mediated ROS production and AS-induced apoptotic and autophagic cell death. Furthermore, AS induced ROS-mediated inhibition of the PI3K/AKT/mTOR signaling pathway. AS reduced the tumor burden in GBM8401-xenografted nude mice and significantly modulated tumor xenografts by inducing anti-EMT, apoptosis, and autophagy. AS could be a potential antitumor agent in human glioblastoma treatment.
背景
红缘拟层孔菌（AS）对多种癌症具有抗癌活性。
目的
本研究旨在探讨 AS 对体外和体内人神经胶质瘤（GBM8401 和 U87MG）细胞的抗癌活性，并解释其潜在的分子机制。
方法
采用 MTT、集落形成、迁移/侵袭实验、免疫印迹、免疫荧光、TUNEL、Annexin V/PI 染色、AO 染色、GFP-LC3 转染、TEM、qPCR、siLC3、DCFH2-DA 测定、以及裸鼠异种移植等方法评估 AS 治疗的潜力。
结果
AS 处理可延缓神经胶质瘤细胞的生长并抑制集落形成。AS 通过抑制侵袭和迁移、增加 E-钙黏蛋白表达、降低 Twist、Snail 和 N-钙黏蛋白表达、抑制 Wnt/β-连环蛋白通路，减弱 EMT 表型，从而抑制 GBM8401 和 U87MG 细胞的迁移和侵袭。此外，AS 通过激活 caspase-3、裂解 PARP 以及上调 Bax 和下调 Bcl-2，在两种细胞系中诱导细胞凋亡。TUNEL 实验和 Annexin V/PI 染色表明 AS 诱导晚期细胞凋亡。有趣的是，AS 通过 LC3-II 积累、AVO 形成、GFP-LC3 斑点形成、p62/SQSTM1 表达和 ATG4B 抑制诱导自噬细胞死亡，在 GBM8401 和 U87MG 细胞中诱导自噬。TEM 数据显示，AS 有利于自噬体和自噬溶酶体的形成。自噬抑制剂 3-MA/CQ 和 LC3 敲低抑制 AS 诱导的神经胶质瘤细胞凋亡，表明自噬抑制降低了 AS 诱导的凋亡。值得注意的是，抗氧化剂 N-乙酰半胱氨酸（NAC）抑制了 AS 诱导的 ROS 产生和 AS 诱导的细胞凋亡和自噬。此外，AS 诱导的 ROS 抑制了 PI3K/AKT/mTOR 信号通路。AS 在 GBM8401 裸鼠异种移植模型中减少了肿瘤负担，并通过诱导抗 EMT、凋亡和自噬显著调节肿瘤异种移植物。AS 可能成为人类神经胶质瘤治疗的一种潜在的抗肿瘤药物。
Zotero 插件