Department of Chemistry 'Ugo Schiff', University of Florence, via della Lastruccia 3-13, Sesto Fiorentino, 50019, Firenze, Italy.
Department of Chemical Sciences, University of Naples Federico II, Naples, Italy.
Cancer Chemother Pharmacol. 2022 Jun;89(6):809-823. doi: 10.1007/s00280-022-04438-y. Epub 2022 May 11.
Ovarian cancer is the fifth leading cause of cancer-related deaths in women. Standard treatment consists of tumor debulking surgery followed by platinum and paclitaxel chemotherapy; yet, despite the initial response, about 70-75% of patients develop resistance to chemotherapy. Gold compounds represent a family of very promising anticancer drugs. Among them, we previously investigated the cytotoxic and pro-apoptotic properties of Au(NHC) and Au(NHC)PF, i.e., a monocarbene gold(I) complex and the corresponding bis(carbene) complex. Gold compounds are known to alter the redox state of cells interacting with free cysteine and selenocysteine residues of several proteins. Herein, a redox proteomic study has been carried out to elucidate the mechanisms of cytotoxicity in A2780 human ovarian cancer cells.
A biotinylated iodoacetamide labeling method coupled with mass spectrometry was used to identify oxidation-sensitive protein cysteines.
Gold carbene complexes cause extensive oxidation of several cellular proteins; many affected proteins belong to two major functional classes: carbohydrate metabolism, and cytoskeleton organization/cell adhesion. Among the affected proteins, Glyceraldehyde-3-phosphate dehydrogenase inhibition was proved by enzymatic assays and by ESI-MS studies. We also found that Au(NHC)PF inhibits mitochondrial respiration impairing complex I function. Concerning the oxidized cytoskeletal proteins, gold binding to the free cysteines of actin was demonstrated by ESI-MS analysis. Notably, both gold compounds affected cell migration and invasion.
In this study, we deepened the mode of action of Au(NHC) and Au(NHC)PF, identifying common cellular targets but confirming their different influence on the mitochondrial function.
卵巢癌是女性癌症相关死亡的第五大主要原因。标准治疗包括肿瘤减瘤手术，随后进行铂类和紫杉醇化疗；然而，尽管最初有反应，但约 70-75%的患者对化疗产生耐药性。金化合物代表了一类非常有前途的抗癌药物。在它们之中，我们之前研究了 Au(NHC) 和 Au(NHC)PF 的细胞毒性和促凋亡特性，即单卡宾金（I）配合物和相应的双（卡宾）配合物。金化合物已知通过与几种蛋白质的游离半胱氨酸和硒代半胱氨酸残基相互作用来改变细胞的氧化还原状态。在此，进行了氧化还原蛋白质组学研究，以阐明 A2780 人卵巢癌细胞中细胞毒性的机制。
使用生物素化碘乙酰胺标记方法结合质谱法来鉴定氧化敏感的蛋白质半胱氨酸。
金卡宾配合物导致几种细胞蛋白的广泛氧化；许多受影响的蛋白质属于两个主要功能类别：碳水化合物代谢和细胞骨架组织/细胞黏附。在受影响的蛋白质中，通过酶促测定和 ESI-MS 研究证明了甘油醛-3-磷酸脱氢酶的抑制作用。我们还发现 Au(NHC)PF 抑制线粒体呼吸，损害复合物 I 功能。关于氧化的细胞骨架蛋白，通过 ESI-MS 分析证明了金与肌动蛋白的游离半胱氨酸结合。值得注意的是，两种金化合物都影响细胞迁移和侵袭。
在这项研究中，我们深入研究了 Au(NHC) 和 Au(NHC)PF 的作用机制，确定了常见的细胞靶标，但证实了它们对线粒体功能的不同影响。
