Department of Chemical Sciences, University of Naples Federico II, via Cintia 21, I-80126 Napoli, Italy.
NanoHybrid BioInterfaces Laboratory (NHBIL), Department of Chemical Sciences, University of Catania, viale Andrea Doria, 6, 95125 Catania, Italy.
Dalton Trans. 2023 Jul 4;52(26):9058-9067. doi: 10.1039/d3dt01517c.
Cisplatin (CisPt), a platinum-based chemotherapeutic widely used in the treatment of various cancers, has multiple mechanisms of action, including nuclear DNA (nDNA) and mitochondrial DNA (mDNA) damage and cytoskeletal perturbations affecting, in turn, the membrane transporter activity. CisPt binding to proteins and enzymes may modulate its biochemical mechanism of action and is associated with cancer cell resistance to the drug. In this work, we investigate the interaction between cisplatin and angiogenin (Ang), a protein strongly expressed in many types of cancer and a potent angiogenic factor. The adduct formed upon reaction of CisPt with Ang (Ang@CisPt) was characterized by X-ray crystallography to evidence the exact platination site and by UV-visible (UV-vis) absorption and circular dichroism (CD) spectroscopies to shed light on any possible change in the protein conformation. Furthermore, high-resolution electrospray ionization (ESI) mass spectrometry was utilized to evaluate the Ang : CisPt stoichiometry of the Ang@CisPt adduct. The effect of the Ang@CisPt adduct on a prostate cancer cell line (PC-3) was tested by colorimetric assays in terms of cell viability, at both levels of nuclear and mitochondrial damage, and reactive oxygen species (ROS) production. Cellular imaging by laser scanning confocal microscopy (LSM) was utilized to scrutinize the cytoskeleton actin reorganization and the lysosome and mitochondria organelle perturbation. These studies highlight the possibility of new molecular pathways and targets for CisPt activity.
顺铂（CisPt）是一种广泛用于治疗各种癌症的铂类化疗药物，具有多种作用机制，包括核 DNA（nDNA）和线粒体 DNA（mDNA）损伤以及影响细胞骨架的细胞骨架扰动，进而影响膜转运体活性。 CisPt 与蛋白质和酶的结合可能调节其生化作用机制，并与癌细胞对药物的耐药性有关。在这项工作中，我们研究了顺铂与血管生成素（Ang）之间的相互作用，Ang 是一种在许多类型的癌症中强烈表达的蛋白质，也是一种有效的血管生成因子。通过 X 射线晶体学对 CisPt 与 Ang 反应形成的加合物（Ang@CisPt）进行了表征，以证明确切的铂化位点，并通过紫外可见（UV-vis）吸收和圆二色性（CD）光谱研究任何可能的蛋白质构象变化。此外，还利用高分辨率电喷雾电离（ESI）质谱评估了 Ang@CisPt 加合物的 Ang：CisPt 化学计量比。通过比色法测定 Ang@CisPt 加合物对前列腺癌细胞系（PC-3）的影响，评估其对核和线粒体损伤水平以及活性氧（ROS）产生的细胞活力的影响。通过激光扫描共聚焦显微镜（LSM）进行细胞成像，研究细胞骨架肌动蛋白重排以及溶酶体和线粒体细胞器扰动。这些研究强调了 CisPt 活性的新分子途径和靶标的可能性。
