College of Applied Science and Technology, Beijing Key Laboratory of Bioactive Substances and Functional Foods, Beijing Union University, Beijing 100101, China.
Beijing National Laboratory for Molecular Sciences, CAS Research/Education Centre for Excellence in Molecular Sciences, National Centre for Mass Spectrometry in Beijing, CAS Key Laboratory of Analytical Chemistry for Living Biosystems, Institute of Chemistry, Chinese Academy of Sciences, Beijing 100190, China.
Int J Mol Sci. 2023 Apr 24;24(9):7756. doi: 10.3390/ijms24097756.
The ruthenium polypyridine complex [Ru(dppa)(pytp)] (PF) (termed as ZQX-1), where dppa = 4,7-diphenyl-1,10-phenanthroline and pytp = 4'-pyrene-2,2':6',2''-terpyridine, has been shown a high and selective cytotoxicity to hypoxic and cisplatin-resistant cancer cells either under irradiation with blue light or upon two-photon excitation. The IC values of ZQX-1 towards A549 cancer cells and HEK293 health cells are 0.16 ± 0.09 µM and >100 µM under irradiation at 420 nm, respectively. However, the mechanism of action of ZQX-1 remains unclear. In this work, using the quantitative proteomics method we identified 84 differentially expressed proteins (DEPs) with |fold-change| ≥ 1.2 in A549 cancer cells exposed to ZQX-1 under irradiation at 420 nm. Bioinformatics analysis of the DEPs revealed that photoactivated ZQX-1 generated reactive oxygen species (ROS) to activate oxidative phosphorylation signaling to overproduce ATP; it also released ROS and pyrene derivative to damage DNA and arrest A549 cells at S-phase, which synergistically led to oncotic necrosis and apoptosis of A549 cells to deplete excess ATP, evidenced by the elevated level of PRAP1 and cleaved capase-3. Moreover, the DNA damage inhibited the expression of DNA repair-related proteins, such as RBX1 and GPS1, enhancing photocytotoxicity of ZQX-1, which was reflected in the inhibition of integrin signaling and disruption of ribosome assembly. Importantly, the photoactivated ZQX-1 was shown to activate hypoxia-inducible factor 1A (HIF1A) survival signaling, implying that combining use of ZQX-1 with HIF1A signaling inhibitors may further promote the photocytotoxicity of the prodrug.
钌多吡啶配合物[Ru(dppa)(pytp)]（PF）（称为 ZQX-1），其中 dppa = 4,7-二苯基-1,10-菲咯啉，pytp = 4'-并噻吩-2,2':6',2''-三联吡啶，已显示出对缺氧和顺铂耐药癌细胞具有高选择性细胞毒性，无论是在蓝光照射下还是在双光子激发下。在 420nm 照射下，ZQX-1 对 A549 癌细胞和 HEK293 健康细胞的 IC 值分别为 0.16±0.09µM 和 >100µM。然而，ZQX-1 的作用机制仍不清楚。在这项工作中，我们使用定量蛋白质组学方法，在 420nm 光照下，鉴定出暴露于 ZQX-1 的 A549 癌细胞中 84 个差异表达蛋白（DEPs），|fold-change|≥1.2。对 DEPs 的生物信息学分析表明，光激活的 ZQX-1 产生活性氧（ROS）来激活氧化磷酸化信号以过度产生 ATP；它还释放 ROS 和并噻吩衍生物来损伤 DNA 并将 A549 细胞阻滞在 S 期，这协同导致 A549 细胞的胀亡性坏死和凋亡，从而消耗过多的 ATP，这一点可从 PRAP1 和 cleaved capase-3 的水平升高得到证明。此外，DNA 损伤抑制了 DNA 修复相关蛋白的表达，如 RBX1 和 GPS1，增强了 ZQX-1 的光细胞毒性，这反映在整合素信号的抑制和核糖体组装的破坏上。重要的是，光激活的 ZQX-1 被证明激活了缺氧诱导因子 1A（HIF1A）存活信号，这意味着将 ZQX-1 与 HIF1A 信号抑制剂联合使用可能进一步促进前药的光细胞毒性。
