Abstract：
BACKGROUND: Breast cancer bone metastasis is closely associated with the bone microenvironment. Zuogui Pill (ZGP), a clinically approved formulation in China, effectively regulates the bone microenvironment for the prevention and treatment of osteoporosis.
OBJECTIVE: Few reports have utilized the ZGP for bone metastasis models. This study investigated the intervention and bone-protective properties of ZGP against breast cancer bone metastasis, explored the potential mechanism, and screened for its active compositions by molecules fishing.
METHODS: To investigate the intervention efficacy of ZGP and its protein-level mechanism of action, the mouse bone metastasis model and in vitro cell co-culture model were constructed. Affinity ultrafiltration, molecular docking, cellular thermal shift assay and physical scale detection were used to investigate the affinity components of the RANKL protein in ZGP.
RESULTS: The administration of ZGP combined with zoledronic acid inhibited the development of tumors and secondary lung metastasis in mice. This translated to a prolonged survival period and enhanced quality of life. ZGP could disrupt the malignant cycle by modulating the Piezo1-Notch-1-GPX4 signaling pathway in the \"bone-cancer\" communication in the cell co-culture model. Furthermore, 25 chemical components of ZGP were identified, with 10 active compounds exhibiting significant affinity for the RANKL protein.
CONCLUSIONS: The findings of this work highlighted ZGP\'s potential for intervening in the progression of breast cancer bone metastasis. Thus, this investigation served as an experimental foundation for expanding the application scope of ZGP and for advancing drug development efforts in bone metastasis treatment.
OBJECTIVE: Few reports have utilized the ZGP for bone metastasis models. This study investigated the intervention and bone-protective properties of ZGP against breast cancer bone metastasis, explored the potential mechanism, and screened for its active compositions by molecules fishing.
METHODS: To investigate the intervention efficacy of ZGP and its protein-level mechanism of action, the mouse bone metastasis model and in vitro cell co-culture model were constructed. Affinity ultrafiltration, molecular docking, cellular thermal shift assay and physical scale detection were used to investigate the affinity components of the RANKL protein in ZGP.
RESULTS: The administration of ZGP combined with zoledronic acid inhibited the development of tumors and secondary lung metastasis in mice. This translated to a prolonged survival period and enhanced quality of life. ZGP could disrupt the malignant cycle by modulating the Piezo1-Notch-1-GPX4 signaling pathway in the \"bone-cancer\" communication in the cell co-culture model. Furthermore, 25 chemical components of ZGP were identified, with 10 active compounds exhibiting significant affinity for the RANKL protein.
CONCLUSIONS: The findings of this work highlighted ZGP\'s potential for intervening in the progression of breast cancer bone metastasis. Thus, this investigation served as an experimental foundation for expanding the application scope of ZGP and for advancing drug development efforts in bone metastasis treatment.
摘要：
背景：乳腺癌骨转移与骨微环境密切相关。左归丸（ZGP），在中国临床批准的制剂，有效调节骨微环境,预防和治疗骨质疏松。
目的：很少有报道将ZGP用于骨转移模型。本研究探讨了ZGP对乳腺癌骨转移的干预和骨保护作用。探索潜在的机制，并通过分子捕鱼筛选其活性成分。
方法：探讨ZGP干预效果及其蛋白水平作用机制，建立小鼠骨转移模型和体外细胞共培养模型。亲和超滤，分子对接,采用细胞热转移法和物理尺度检测法研究ZGP中RANKL蛋白的亲和成分。
结果：ZGP联合唑来膦酸对小鼠肿瘤的发展和继发性肺转移具有抑制作用。这意味着延长了生存期并提高了生活质量。ZGP可以通过调节细胞共培养模型中“骨癌”通讯中的Piezo1-Notch-1-GPX4信号通路来破坏恶性周期。此外，确定了ZGP的25种化学成分，具有10种对RANKL蛋白表现出显著亲和力的活性化合物。
结论：这项工作的发现强调了ZGP在干预乳腺癌骨转移进展方面的潜力。因此，这项研究为扩大ZGP的应用范围和推进骨转移治疗药物的开发工作奠定了实验基础.
目的：很少有报道将ZGP用于骨转移模型。本研究探讨了ZGP对乳腺癌骨转移的干预和骨保护作用。探索潜在的机制，并通过分子捕鱼筛选其活性成分。
方法：探讨ZGP干预效果及其蛋白水平作用机制，建立小鼠骨转移模型和体外细胞共培养模型。亲和超滤，分子对接,采用细胞热转移法和物理尺度检测法研究ZGP中RANKL蛋白的亲和成分。
结果：ZGP联合唑来膦酸对小鼠肿瘤的发展和继发性肺转移具有抑制作用。这意味着延长了生存期并提高了生活质量。ZGP可以通过调节细胞共培养模型中“骨癌”通讯中的Piezo1-Notch-1-GPX4信号通路来破坏恶性周期。此外，确定了ZGP的25种化学成分，具有10种对RANKL蛋白表现出显著亲和力的活性化合物。
结论：这项工作的发现强调了ZGP在干预乳腺癌骨转移进展方面的潜力。因此，这项研究为扩大ZGP的应用范围和推进骨转移治疗药物的开发工作奠定了实验基础.
