Saikosaponin d通过干扰NQO1介导的细胞内氧化还原平衡来增强阿霉素对药物抗药性乳腺癌的抗肿瘤作用
Saikosaponin D potentiates the antineoplastic effects of doxorubicin in drug-resistant breast cancer through perturbing NQO1-mediated intracellular redox balance
影响因子:8.30000
分区:医学1区 Top / 药物化学1区 全科医学与补充医学1区 药学1区 植物科学1区
发表日期:2024 Oct
作者:
Fazhen Luo, Juan Yang, Xiuru Yang, Jinxia Mi, Taiwei Ye, Guowen Li, Yan Xie
摘要
对阿霉素（DOX）的耐药性显着限制了其在乳腺癌（BC）患者中的治疗功效。 Saikosaponin D（SSD）是一种源自传统的Herb radix bupleuri的三萜皂苷，由于其显着的抗肿瘤活性，在临床前研究中表现出了作为化学治疗敏化剂的希望。然而，SSD在耐DOX耐药的BC细胞中的作用和机制仍然在很大程度上没有探索。这项研究旨在研究SSD对耐DOX耐药性BC的化学敏化作用，以及在体外和体外的基本分子机制，包括体外和体内的增长。评估SSD和DOX对抗性BC细胞的协同作用。使用活性氧（ROS），GSH/GSSG，NADPH/NADP+和NADH/NAD+检测来评估SSD对细胞氧化还原稳态的影响。进行了蛋白质印迹，细胞周期分布测定和DOX摄取测定法，以进一步阐明SSD的抗塑性机制。最后，建立了裸鼠中的皮下MCF7/DOX细胞异种移植模型，以鉴定SSD的体内抗癌作用与DOX.SSD相结合，可显着抑制细胞的活力，增殖和克隆形成，并增强DOX的Dox的抗药性抗癌作用。从机械上讲，SSD降低了STAT1，NQO1和PGC-1α蛋白水平，从而导致细胞氧化还原失衡，过度ROS产生以及GSH，NADPH和NADH的耗竭。 SSD通过破坏氧化还原稳态引起DNA损伤，从而导致G0/G1相细胞周期停滞。此外，SSD通过抑制P-GP蛋白表达和外排活性增加了BC细胞中DOX的积累。我们首次证明SSD通过灭绝STAT1/NQO1/PGC-1α信号通路来破坏细胞氧化还原稳态，从而增强了化学耐药性BC细胞对DOX的敏感性。这项研究为抗药性BC治疗中的SSD作为辅助药提供了证据。
Abstract
Drug resistance to doxorubicin (DOX) significantly limits its therapeutic efficacy in breast cancer (BC) patients. Saikosaponin D (SSD), a triterpene saponin derived from the traditional herb Radix Bupleuri, has shown promise as a chemotherapeutic sensitizer in preclinical studies due to its notable antitumor activity. However, the role and mechanism of SSD in DOX-resistant BC cells remain largely unexplored.This study aimed to investigate the chemosensitizing effect of SSD on DOX-resistant BC and the underlying molecular mechanisms both in vitro and in vivo.In vitro assays, including cell viability, clone formation, three-dimensional tumor spheroid growth, and apoptosis analysis, were conducted to evaluate the synergistic effect of SSD and DOX on resistant BC cells. Reactive oxygen species (ROS), GSH/GSSG, NADPH/NADP+, and NADH/NAD+ detections were employed to assess the impact of SSD on cellular redox homeostasis. Western blotting, cell cycle distribution assay, and DOX uptake assay were performed to further elucidate the possible antineoplastic mechanism of SSD. Finally, a subcutaneous MCF7/DOX cell xenografted model in nude mice was established to identify the in vivo anticarcinogenic effect of SSD combined with DOX.SSD significantly inhibited cell viability, proliferation, and clone formation, enhancing DOX's anticancer efficacy in vitro and in vivo. Mechanistically, SSD reduced STAT1, NQO1, and PGC-1α protein levels, leading to cellular redox imbalance, excessive ROS generation, and depletion of GSH, NADPH, and NADH. SSD induced DNA damage by disrupting redox homeostasis, resulting in G0/G1 phase cell cycle arrest. Additionally, SSD increased DOX accumulation in BC cells via inhibiting P-gp protein expression and efflux activity.We demonstrated for the first time that SSD enhances the sensitivity of chemoresistant BC cells to DOX by disrupting cellular redox homeostasis through inactivation of the STAT1/NQO1/PGC-1α signaling pathway. This study provides evidence for SSD as an adjuvant agent in drug-resistant BC treatment.