Ouyang Jiabao, Li Nan, Liu Chunbo, Zhang Yu
Department of Ultrasound, The First Affiliated Hospital of Harbin Medical University No. 199, Xidazhi Road, Nangang District City Harbin Heilongjiang Province 150001 China
Nanoscale Adv. 2025 Jun 12. doi: 10.1039/d5na00280j.
: Hepatocellular carcinoma (HCC) is one of the primary causes of cancer-associated death worldwide, which has few viable therapeutic options at later stages. Cuproptosis, a newly discovered kind of programmed cell death, gives a new therapeutic target for HCC. Nanobubble (NB)-mediated sonodynamic therapy (SDT) has been a prospective method for augmenting the efficacy and delivery of therapeutic agents. This study investigates the potential of NB-mediated SDT to enhance cuproptosis in HCC treatment, aiming to evaluate the efficacy of NB-mediated SDT in enhancing cuproptosis in HCC cells and to clarify the potential mechanisms. : NBs were engineered to encapsulate the sonosensitizer protoporphyrin IX (PPIX) and the cuproptosis inducer elesclomol-Cu (ES-Cu), forming PPIX@ES-Cu NBs, a targeted delivery system activated by ultrasound. HCC cell lines (Hepa1-6) were treated with these PPIX@ES-Cu NBs followed by low-intensity ultrasound (LIUS) exposure. Mitochondrial function and cell viability were evaluated through CCK assays and fluorescence microscopy. qPCR was implemented for examining the expression of cuproptosis-associated genes. : The NB-mediated SDT significantly enhanced the delivery of copper ions and sonosensitizers to HCC cells. Upon ultrasound activation, there was a substantial increase in ROS production and intracellular copper accumulation. This led to mitochondrial dysfunction, characterized by decreased mitochondrial membrane potential and disrupted tricarboxylic acid (TCA) cycle enzyme activities. Treated cells exhibited increased expression of cuproptosis markers, including upregulation of lipoylated mitochondrial proteins and proteotoxic stress indicators. Cell viability assays demonstrated a considerable decrease in HCC cell survival compared to controls. : NB-mediated SDT effectively enhances cuproptosis in HCC cells by facilitating targeted delivery and activation of therapeutic agents. The synergistic effect of ROS generation and copper accumulation induces cuproptosis, offering a novel and potent therapeutic strategy for HCC treatment. These results lay the groundwork for more research and possible clinical use of this combo treatment.
肝细胞癌（HCC）是全球癌症相关死亡的主要原因之一，在晚期阶段几乎没有可行的治疗选择。铜死亡是一种新发现的程序性细胞死亡，为HCC提供了一个新的治疗靶点。纳米气泡（NB）介导的声动力疗法（SDT）一直是一种增强治疗剂疗效和递送的前瞻性方法。本研究探讨NB介导的SDT在HCC治疗中增强铜死亡的潜力，旨在评估NB介导的SDT增强HCC细胞铜死亡的疗效，并阐明其潜在机制。：将NB设计成包裹声敏剂原卟啉IX（PPIX）和铜死亡诱导剂依斯氯铵铜（ES-Cu），形成PPIX@ES-Cu NB，这是一种由超声激活的靶向递送系统。用这些PPIX@ES-Cu NB处理HCC细胞系（Hepa1-6），然后进行低强度超声（LIUS）照射。通过CCK试验和荧光显微镜评估线粒体功能和细胞活力。采用qPCR检测铜死亡相关基因的表达。：NB介导的SDT显著增强了铜离子和声敏剂向HCC细胞的递送。超声激活后，活性氧生成和细胞内铜积累显著增加。这导致线粒体功能障碍，其特征是线粒体膜电位降低和三羧酸（TCA）循环酶活性受到破坏。处理后的细胞显示铜死亡标志物的表达增加，包括脂酰化线粒体蛋白和蛋白毒性应激指标的上调。细胞活力试验表明，与对照组相比，HCC细胞存活率显著降低。：NB介导的SDT通过促进治疗剂的靶向递送和激活，有效增强HCC细胞中的铜死亡。活性氧生成和铜积累的协同作用诱导铜死亡，为HCC治疗提供了一种新的有效治疗策略。这些结果为这种联合治疗的更多研究和可能的临床应用奠定了基础。
