铜负载葡萄糖氧化酶纳米药物用于肝癌的化学动力学治疗

The Copper-Loaded Glucose Oxidase Nanomedicines for Chemodynamic Therapy of Hepatocellular Carcinoma

摘要: 目的：为了达到靶向肿瘤部位实现肝癌的精准治疗，设计合成一种负载葡萄糖氧化酶(GOx)和铜离子(Cu² )的纳米药物(Cu@GoxNM)，通过化学动力学治疗达到对肿瘤细胞的杀伤作用。方法：通过“一锅法”合成铜负载葡萄糖氧化酶的纳米药物，通过扫描电镜和元素映射检测相关元素来验证纳米药物的合成。验证该纳米药物的级联催化产生活性氧作用。通过MTT细胞毒活实验，细胞活性氧的检测和活死细胞染色实验来验证其对细胞的影响，由此来证明化学动力学治疗的效果。结果：成功合成了铜负载葡萄糖氧化酶的纳米药物Cu@GoxNM；Cu@GoxNM对HUH7细胞的促凋亡作用与其浓度呈正相关；与对照细胞相比Cu@GoxNM致HUH7细胞产生更多的活性氧并发生细胞凋亡。结论：Cu@GoxNM纳米药物能明显促进肝癌细胞的凋亡，其是通过产生过量的活性氧达到化学动力学疗法作用。

Abstract: Objective: In order to achieve precise treatment of liver cancer by targeting the tumour site, to design and synthesize a nanomedicine (Cu@GoxNM) loaded with glucose oxidase (GOx) and copper ions (Cu² ), which can kill the tumour cells through chemodynamic therapy. Methods: Synthesis of copper-loaded glucose oxidase nanomedicine by “one-pot method” was demonstrated by scanning electron microscopy (SEM) and elemental mapping for the detection of relevant elements. The cascade-catalyzed generation of reactive oxygen species by the nanomedicine was experimentally verified. The effects of the nanomedicine on cells were verified by MTT cytotoxicity assay, cellular reactive oxygen species assay, and staining assay of live and dead cells to demonstrate the effectiveness of chemodynamic therapy. Results: The copper-loaded glucose oxidase nanomedicine Cu@GoxNM was successfully synthesised; the pro-apoptotic effect of Cu@GoxNM on HUH7 cells was positively correlated with its concentration; Cu@GoxNM resulted in the production of more reactive oxygen species and apoptosis in HUH7 cells compared to control cells. Conclusion: Cu@GoxNM nanomedicine significantly promotes the apoptosis of hepatocellular carcinoma cells, and it might be the effect of chemodynamic therapy by generating excessive reactive oxygen species.