Research Center for Health and Nutrition, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, China; Pharmacy Department, Shanghai Integrated Traditional Chinese and Western Medicine Hospital, 184 Baoding Road, Shanghai 200082, China.
Research Center for Health and Nutrition, Shanghai University of Traditional Chinese Medicine, 1200 Cailun Road, Shanghai 201203, China.
Phytomedicine. 2024 Oct;133:155945. doi: 10.1016/j.phymed.2024.155945. Epub 2024 Aug 8.
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.
多柔比星（DOX）耐药性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）是一种从传统草药柴胡中提取的三萜皂苷，由于其显著的抗肿瘤活性，在临床前研究中已显示出作为化疗增敏剂的潜力。然而，SSD 在 DOX 耐药 BC 细胞中的作用和机制在很大程度上仍未得到探索。
本研究旨在探讨 SSD 对 DOX 耐药 BC 的化疗增敏作用及其在体内外的潜在分子机制。
采用细胞活力、克隆形成、三维肿瘤球体生长和凋亡分析等体外实验评估 SSD 和 DOX 对耐药 BC 细胞的协同作用。采用活性氧（ROS）、GSH/GSSG、NADPH/NADP 和 NADH/NAD 检测评估 SSD 对细胞氧化还原平衡的影响。通过 Western blot、细胞周期分布分析和 DOX 摄取实验进一步阐明 SSD 的可能抗肿瘤机制。最后，建立裸鼠皮下 MCF7/DOX 细胞异种移植模型，以确定 SSD 联合 DOX 的体内抗癌作用。
SSD 显著抑制细胞活力、增殖和克隆形成，增强了 DOX 在体内外的抗癌疗效。机制上，SSD 降低了 STAT1、NQO1 和 PGC-1α 蛋白水平，导致细胞内氧化还原失衡、ROS 生成过多、GSH、NADPH 和 NADH 耗竭。SSD 通过破坏氧化还原平衡诱导 DNA 损伤，导致 G0/G1 期细胞周期停滞。此外，SSD 通过抑制 P-gp 蛋白表达和外排活性增加 BC 细胞内 DOX 的积累。
我们首次证明，通过失活 STAT1/NQO1/PGC-1α 信号通路破坏细胞氧化还原平衡，SSD 增强了耐药 BC 细胞对 DOX 的敏感性。这项研究为 SSD 作为耐药性 BC 治疗的辅助药物提供了证据。
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