Zhu Yuxuan, Huang Zhichao, Tian Shichao, Wang Qi, Wu Fan, Liu Jingwen, Wang Ping, Wan Hao, Zhuang Liujing, Jiang Deming
Biosensor National Special Laboratory, Key Laboratory for Biomedical Engineering of Education Ministry, Department of Biomedical Engineering, Zhejiang University, Hangzhou, 310027, China.
State Key Laboratory of Transducer Technology, Chinese Academy of Sciences, Shanghai, 200050, China.
Microsyst Nanoeng. 2025 Aug 14;11(1):154. doi: 10.1038/s41378-025-01016-z.
In cancer cells, higher reactive oxygen species (ROS) than normal cells were observed due to hypermetabolism. The redox balance in cancer cells relies on accordingly upregulated antioxidant capacity. By manipulating oxidation and antioxidant systems, chemotherapeutic drugs can selectively kill cancer cells without hurting normal cells. As three-dimensional (3D) in vitro models, such as spheroids and organoids, have become widely used in cancer research, traditional detection methods (e.g., absorption tests or titration) are inadequate for detecting in 3D environments. Thus, it is crucial to find a new method to detect oxidative stress of 3D in vitro cancer models. Here, a nanocomposite electrochemical biosensor was exploited to evaluate oxidative stress of cancer cells cultured in the 3D environment. The oxidation-regulatory capacity of honokiol, a Magnolia genus-derived anti-cancer molecule, was evaluated. A screen-printed electrode (SPCE) was modified with reduced graphene oxide (RGO) and platinum nanoparticles (Pt NPs) to get Pt NPs/RGO/SPCE. Then the gelatin methacrylate/reduced graphene oxide (GelMA/RGO) hydrogel was applied to immobilized NCI-H1975 in a 3D bionic environment to get NCI-H1975/GelMA/RGO/Pt NPs/RGO/SPCE. After optimizing the experiment condition, the Pt NPs/RGO/SPCE showed a detection threshold of 0.65 μM and a linear field from 1 to 10 μM for HO detection while the NCI-H1975/GelMA/RGO/Pt NPs/RGO/SPCE sensitively responded to HO-induced oxidative stress. By utilizing the NCI-H1975/GelMA/RGO/Pt NPs/RGO/SPCE we found honokiol (a natural polyphenol constituent) inhibits NCI-H1975 by inducing oxidative stress. This simple cell-based electrochemical biosensor can in situ evaluate oxidative stress of 3D cancer models conveniently. It can also be easily extended to the study of the mechanism of action of other drugs and holds broad application prospects in the fields of new drug development and drug repurposing.
在癌细胞中，由于代谢亢进，观察到其活性氧（ROS）水平高于正常细胞。癌细胞中的氧化还原平衡相应地依赖于上调的抗氧化能力。通过操纵氧化和抗氧化系统，化疗药物可以选择性地杀死癌细胞而不损伤正常细胞。随着三维（3D）体外模型，如球体和类器官，已广泛应用于癌症研究，传统的检测方法（如吸收试验或滴定法）不足以在3D环境中进行检测。因此，找到一种新的方法来检测3D体外癌症模型的氧化应激至关重要。在此，利用一种纳米复合电化学生物传感器来评估在3D环境中培养的癌细胞的氧化应激。评估了厚朴酚（一种木兰属来源的抗癌分子）的氧化调节能力。用还原氧化石墨烯（RGO）和铂纳米颗粒（Pt NPs）修饰丝网印刷电极（SPCE），得到Pt NPs/RGO/SPCE。然后将明胶甲基丙烯酸酯/还原氧化石墨烯（GelMA/RGO）水凝胶应用于在3D仿生环境中固定NCI-H1975，得到NCI-H1975/GelMA/RGO/Pt NPs/RGO/SPCE。优化实验条件后，Pt NPs/RGO/SPCE对HO的检测阈值为0.65μM，线性范围为1至10μM，而NCI-H1975/GelMA/RGO/Pt NPs/RGO/SPCE对HO诱导的氧化应激有敏感响应。通过使用NCI-H1975/GelMA/RGO/Pt NPs/RGO/SPCE，我们发现厚朴酚（一种天然多酚成分）通过诱导氧化应激抑制NCI-H1975。这种基于细胞的简单电化学生物传感器可以方便地原位评估3D癌症模型的氧化应激。它还可以很容易地扩展到其他药物作用机制的研究，在新药开发和药物再利用领域具有广阔的应用前景。
