Department of Urology, Department of Science & Technology, The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Quzhou, Zhejiang 324000, PR China.
Department of Urology, Department of Science & Technology, The Quzhou Affiliated Hospital of Wenzhou Medical University, Quzhou People's Hospital, Quzhou, Zhejiang 324000, PR China.
Biomed Pharmacother. 2023 Feb;158:114154. doi: 10.1016/j.biopha.2022.114154. Epub 2022 Dec 28.
Bladder cancer (BC) is a common urological malignancy that still lacks effective treatments. Abietic acid (AA) is an abietane diterpene that possesses various biological activities, including antitumor activity. This study aimed at evaluating the effects of AA on BC cells.
The 3-(4, 5-dimethylthiazolyl-2)-2, 5-diphenyltetrazolium bromide (MTT) assay was used to assess the effects of AA on the viability of BC cells. Annexin-V and FITC staining was used to assess cellular death. The type of cell death was determined by the administration of various specific cell death inhibitors. Commercial kits were used to measure the levels of reactive oxygen species (ROS), intracellular iron, malondialdehyde (MDA), and glutathione (GSH). Real-time polymerase chain reaction (RT-PCR) and western blot analysis were used to assay mRNA and protein levels, respectively. The role of glutathione peroxidase 4 (GPX4) in the antitumor effects of AA was evaluated using the forced expression of GPX4 in BC cells. The impact of HO-1 on the antitumor effects of AA was examined by gene silencing and pharmacological inhibition of the protein. Finally, the antitumor effects of AA were evaluated in xenograft models.
AA selectively inhibited the viability of BC cells but not normal cells. AA-induced ferroptosis in BC cells was evidenced by the upregulation of ROS, intracellular iron, and MDA. AA treatment led to the downregulation of GPX4 and the upregulation of HO-1 in BC cells. Forced expression of GPX4 or inhibition of HO-1 resulted in decreased ferroptosis triggered by AA in BC cells. AA also showed synergistic effects with various chemotherapeutic agents against BC and inhibited the growth of BC cells in vivo.
This study revealed AA-induced ferroptosis in BC cells both in vitro and in vivo. AA might be applied as a promising agent for the treatment of BC.
膀胱癌（BC）是一种常见的泌尿系统恶性肿瘤，目前仍缺乏有效的治疗方法。枞酸（AA）是一种枞烷二萜，具有多种生物活性，包括抗肿瘤活性。本研究旨在评估 AA 对 BC 细胞的影响。
采用 3-(4,5-二甲基噻唑基-2)-2,5-二苯基四氮唑溴盐（MTT）法评估 AA 对 BC 细胞活力的影响。采用 Annexin-V 和 FITC 染色评估细胞死亡。通过给予各种特定的细胞死亡抑制剂来确定细胞死亡的类型。采用商业试剂盒测定活性氧（ROS）、细胞内铁、丙二醛（MDA）和谷胱甘肽（GSH）的水平。实时聚合酶链反应（RT-PCR）和蛋白质印迹分析分别用于检测 mRNA 和蛋白水平。通过在 BC 细胞中强制表达 GPX4 来评估谷胱甘肽过氧化物酶 4（GPX4）在 AA 抗肿瘤作用中的作用。通过基因沉默和蛋白药理学抑制来研究 HO-1 对 AA 抗肿瘤作用的影响。最后，在异种移植模型中评估 AA 的抗肿瘤作用。
AA 选择性抑制 BC 细胞而不影响正常细胞的活力。AA 诱导 BC 细胞发生铁死亡，表现为 ROS、细胞内铁和 MDA 的上调。AA 处理导致 BC 细胞中 GPX4 下调和 HO-1 上调。在 BC 细胞中强制表达 GPX4 或抑制 HO-1 可减少 AA 触发的铁死亡。AA 还与各种化疗药物对 BC 的协同作用，并抑制 BC 细胞在体内的生长。
本研究揭示了 AA 诱导的 BC 细胞在体外和体内的铁死亡。AA 可能被应用为治疗 BC 的有前途的药物。
Zotero 插件