柴胡皂苷D增强多柔比星在耐药乳腺癌中的抗肿瘤作用，通过扰乱NQO1介导的细胞内氧化还原平衡

Saikosaponin D potentiates the antineoplastic effects of doxorubicin in drug-resistant breast cancer through perturbing NQO1-mediated intracellular redox balance

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PHYTOMEDICINE

影响因子:8.3

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

DOI: 10.1016/j.phymed.2024.155945

摘要

多柔比星（DOX）耐药显著限制了其在乳腺癌（BC）患者中的治疗效果。柴胡皂苷D（SSD），一种来源于中药柴胡的三萜皂苷，在临床前研究中显示出作为化疗敏感剂的潜力，因其显著的抗肿瘤活性。然而，SSD在DOX耐药性乳腺癌细胞中的作用及其机制尚未得到充分研究。本研究旨在探讨SSD对耐药乳腺癌的化学敏感作用及其分子机制，包括体外的细胞存活率、克隆形成、三维肿瘤球体生长及凋亡分析。采用ROS、GSH/GSSG、NADPH/NADP+及NADH/NAD+检测评估SSD对细胞氧化还原稳态的影响，Western blot、细胞周期分析及DOX摄取检测进一步阐明其抗肿瘤机制。在裸鼠异种移植模型中验证SSD联合DOX的抗癌效果。结果显示，SSD显著抑制细胞活性、增殖及克隆形成，增强了DOX的抗癌效果。机制方面，SSD通过降低STAT1、NQO1和PGC-1α蛋白水平，导致细胞内氧化还原失衡，产生过量ROS，并耗竭GSH、NADPH和NADH，诱导DNA损伤，造成G0/G1期细胞周期阻滞，并通过抑制P-糖蛋白（P-gp）表达和活动，增加了乳腺癌细胞中DOX的积累。首次证明，SSD通过抑制STAT1/NQO1/PGC-1α信号通路，干扰细胞氧化还原稳态，从而增强耐药乳腺癌细胞对DOX的敏感性，为SSD作为耐药乳腺癌辅助药物提供了理论依据。

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.