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<p>DNA-templated Cu/Pt nanoclusters (DNA-Cu/Pt NCs) were fabricated using four single-stranded oligonucleotides (C14, G14, A14, and T14) as the templates. Their oxidoreductase-like properties, including peroxidase (POD), oxidase (OXD), catalase (CAT), and superoxide dismutase (SOD) activities, were systematically investigated for the first time. The results revealed that all as-prepared nanomaterials exhibited four distinct enzyme-like activities, and both DNA sequences and the molar ratio of precursor metal ions exerted significant impacts on their catalytic performance. When the molar ratio of precursor Cu (Ⅱ) to Pt (Ⅱ) was 10:4, C14-Cu/Pt NCs exhibited superior POD- and CAT-like activities, whereas A14-Cu/Pt NCs showed the strongest OXD-like catalytic performance. On the contrary, A14-Cu NCs possessed the highest SOD-like activity. All three NCs displayed uniform size distribution, and Pt(0)/Pt(Ⅱ) (for bimetallic NCs) as well as Cu(0)/Cu(Ⅰ)/Cu(II) species coexisted on the NC surface. Furthermore, both C14-Cu/Pt NCs and A14-Cu/Pt NCs exhibited excellent glutathione (GSH) oxidase (GSHOx)- and gluocose oxidase (GOx)-mimicking activities. Notably, compared to natural horseradish peroxidase, C14-Cu/Pt NCs exhibited 2.4- and 92.5-fold lower Michaelis constant (<i>K</i><sub>m</sub>) values for the substrates TMB and H<sub>2</sub>O<sub>2</sub>, respectively. Moreover, C14-Cu/Pt NCs displayed strong affinity to H<sub>2</sub>O<sub>2</sub> in CAT-like activity, and <i>K</i><sub>m</sub> value was 0.029 mM. Relative to A14‑Cu/Pt NCs, C14-Cu/Pt NCs also showed a 1.5-fold lower <i>K</i><sub>m</sub> value and a 1.6-fold higher maximum reaction rate (<i>V</i><sub>max</sub>) for GSH. The cascade catalytic reaction involving GOx/GSHox/POD/OXD/CAT/SOD enzyme activities triggered a reactive oxygen species (•OH and <sup>1</sup>O<sub>2</sub>) burst and blocked energy supply in cancer cells. C14-Cu/Pt NCs exhibited good stability in buffer and 10% FBS medium. In vitro experiments confirmed that C14-Cu/Pt NCs could internalized the cells and exhibited good biocompatibility, with low toxicity towards normal 7702 liver cells but remarkable cytotoxicity against HeLa and A549 cancer cells. This work provides a promising strategy for the construction of tumor microenvironment (TME)-responsive multienzyme-like nanozymes for efficient chemodynamic therapy.</p> Graphical abstract <p></p>

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Augmenting in vitro cancer cell toxicity via reactive oxygen species burst and glucose depletion induced by C14-Cu/Pt bimetallic nanozyme cascade reaction

  • 期刊论文

  • 发表日期：

    2026年7月3日

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  • Miao Hu,
  • Chunying Wei

摘要

DNA-templated Cu/Pt nanoclusters (DNA-Cu/Pt NCs) were fabricated using four single-stranded oligonucleotides (C14, G14, A14, and T14) as the templates. Their oxidoreductase-like properties, including peroxidase (POD), oxidase (OXD), catalase (CAT), and superoxide dismutase (SOD) activities, were systematically investigated for the first time. The results revealed that all as-prepared nanomaterials exhibited four distinct enzyme-like activities, and both DNA sequences and the molar ratio of precursor metal ions exerted significant impacts on their catalytic performance. When the molar ratio of precursor Cu (Ⅱ) to Pt (Ⅱ) was 10:4, C14-Cu/Pt NCs exhibited superior POD- and CAT-like activities, whereas A14-Cu/Pt NCs showed the strongest OXD-like catalytic performance. On the contrary, A14-Cu NCs possessed the highest SOD-like activity. All three NCs displayed uniform size distribution, and Pt(0)/Pt(Ⅱ) (for bimetallic NCs) as well as Cu(0)/Cu(Ⅰ)/Cu(II) species coexisted on the NC surface. Furthermore, both C14-Cu/Pt NCs and A14-Cu/Pt NCs exhibited excellent glutathione (GSH) oxidase (GSHOx)- and gluocose oxidase (GOx)-mimicking activities. Notably, compared to natural horseradish peroxidase, C14-Cu/Pt NCs exhibited 2.4- and 92.5-fold lower Michaelis constant (Km) values for the substrates TMB and H2O2, respectively. Moreover, C14-Cu/Pt NCs displayed strong affinity to H2O2 in CAT-like activity, and Km value was 0.029 mM. Relative to A14‑Cu/Pt NCs, C14-Cu/Pt NCs also showed a 1.5-fold lower Km value and a 1.6-fold higher maximum reaction rate (Vmax) for GSH. The cascade catalytic reaction involving GOx/GSHox/POD/OXD/CAT/SOD enzyme activities triggered a reactive oxygen species (•OH and 1O2) burst and blocked energy supply in cancer cells. C14-Cu/Pt NCs exhibited good stability in buffer and 10% FBS medium. In vitro experiments confirmed that C14-Cu/Pt NCs could internalized the cells and exhibited good biocompatibility, with low toxicity towards normal 7702 liver cells but remarkable cytotoxicity against HeLa and A549 cancer cells. This work provides a promising strategy for the construction of tumor microenvironment (TME)-responsive multienzyme-like nanozymes for efficient chemodynamic therapy.

Graphical abstract