Liu Hongli, Wang Luyao, Hu Mengling, Hua Jiale, Lian Xiaofu, Lian Chaoqun, Zhang Jing
Department of Gynecological Oncology, First Affiliated Hospital of Bengbu Medical University, Bengbu, 233030, China.
Department of Genetics, School of Life Sciences, Bengbu Medical University, Bengbu, 233030, China.
Curr Pharm Des. 2025 Aug 19. doi: 10.2174/0113816128383328250808123933.
Ovarian cancer (OC) is a common malignant tumor of the female reproductive system and is usually found at an advanced stage. However, the treatment of OC with conventional the efficacy of surgery and chemotherapy is limited. Brusatol (BRU) is a unique nuclear factor erythroid 2-related factor 2 (Nrf2) pathway inhibitor with significant anti-cancer effects. At the same time, the Nrf2 system also plays a vital role in ferroptosis, which can be used as a new way to treat tumors. This study investigated the mechanism of action of BRU as a novel ferroptosis inducer to inhibit OC cells.
Using bioinformatics to screen for key targets and pathways that act on OC in BRU, and then the effects of BRU on OC cells were examined by cell viability assay, clone formation assay, wound healing assay, and apoptosis assay. The intracellular levels of ROS (Reactive Oxygen Species), Fe2+, glutathione (GSH), and malondialdehyde (MDA) were also quantified. Western blotting analysis was then performed to verify ferroptosis marker proteins and pathways. In addition, the combination of Ferrostatin-1 (Fer-1) and BRU was further tested for ferroptosis-related markers.
By obtaining BRU and OC targets, 171 potential BRU-OC action targets were screened to the core target NQO1. KEGG enrichment analysis showed that the anticancer effects of IBC were mediated through multiple pathways, including the PI3K-AKT and Ras signaling pathways. In vitro results showed that IBC inhibited the proliferation, invasion, and migration of OC cells and induced ferroptosis in OC cells.
We demonstrated that BRU increased intracellular ROS, Fe2+, and MDA levels. It also significantly reduced intracellular GSH level and the expression of two marker proteins for ferroptosis, GPX4 and SLC7A11. Meanwhile, BRU could inhibit the Nrf2/HO-1/NQO1 and AKT/mTOR dual signaling pathways in OC cells. Furthermore, the combination of Ferrostatin-1 (Fer-1) and BRU reversed BRU-induced ferroptosis in OC cells.
In this study, we demonstrated for the first time through bioinformatics, molecular docking technology, and experimental validation that BRU acts as a novel inducer of ferroptosis in ovarian cancer cells by targeting the Nrf2/HO-1/NQO1 and AKT/mTOR dual signaling pathways, and may have great potential in the treatment of ovarian cancer cells.
卵巢癌（OC）是女性生殖系统常见的恶性肿瘤，通常在晚期被发现。然而，传统的手术和化疗对OC的治疗效果有限。布鲁斯他汀（BRU）是一种独特的核因子红细胞2相关因子2（Nrf2）途径抑制剂，具有显著的抗癌作用。同时，Nrf2系统在铁死亡中也起着至关重要的作用，可作为一种新的肿瘤治疗方法。本研究探讨了BRU作为新型铁死亡诱导剂抑制OC细胞的作用机制。
利用生物信息学筛选BRU作用于OC的关键靶点和途径，然后通过细胞活力测定、克隆形成测定、伤口愈合测定和凋亡测定来检测BRU对OC细胞的影响。还对细胞内活性氧（ROS）、Fe2+、谷胱甘肽（GSH）和丙二醛（MDA）水平进行了定量。随后进行蛋白质免疫印迹分析以验证铁死亡标记蛋白和途径。此外，进一步检测了铁抑素-1（Fer-1）与BRU联合使用对铁死亡相关标记物的影响。
通过获取BRU和OC靶点，筛选出171个潜在的BRU-OC作用靶点至核心靶点NQO1。KEGG富集分析表明，BRU的抗癌作用是通过多种途径介导的，包括PI3K-AKT和Ras信号通路。体外实验结果表明，BRU抑制了OC细胞的增殖、侵袭和迁移，并诱导了OC细胞的铁死亡。
我们证明BRU增加了细胞内ROS、Fe2+和MDA水平。它还显著降低了细胞内GSH水平以及铁死亡的两种标记蛋白GPX4和SLC7A11的表达。同时，BRU可抑制OC细胞中的Nrf2/HO-1/NQO1和AKT/mTOR双信号通路。此外，铁抑素-1（Fer-1）与BRU联合使用可逆转BRU诱导的OC细胞铁死亡。
在本研究中，我们首次通过生物信息学、分子对接技术和实验验证表明，BRU通过靶向Nrf2/HO-1/NQO1和AKT/mTOR双信号通路，作为卵巢癌细胞中铁死亡的新型诱导剂，在卵巢癌细胞治疗中可能具有巨大潜力。
