2. Academic Validation
  3. Liquiritin in Aiduqing formula inhibits breast cancer neoangiogenesis by suppressing CTSK-mediated lysosomal degradation of CXCL1 in tumor-associated macrophages

Liquiritin in Aiduqing formula inhibits breast cancer neoangiogenesis by suppressing CTSK-mediated lysosomal degradation of CXCL1 in tumor-associated macrophages

  • Phytomedicine. 2025 Oct 3:148:157358. doi: 10.1016/j.phymed.2025.157358.
Bowen Yang 1,  Jian Luo 1,  Jinyu Wang 2,  Juping Zhang 1,  Yifeng Zheng 1,  Shengqi Wang 3,  Neng Wang 4,  Zhiyu Wang 5
Affiliations

Affiliations

  • 1 State Key Laboratory of Traditional Chinese Medicine Syndrome, Chinese Medicine Guangdong Laboratory, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510006, China; Breast Disease Specialist Hospital of Guangdong Provincial Hospital of Chinese Medicine, the Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China.
  • 2 Breast Disease Specialist Hospital of Guangdong Provincial Hospital of Chinese Medicine, the Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China.
  • 3 State Key Laboratory of Traditional Chinese Medicine Syndrome, Chinese Medicine Guangdong Laboratory, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510006, China; Breast Disease Specialist Hospital of Guangdong Provincial Hospital of Chinese Medicine, the Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, Guangdong Provincial Academy of Chinese Medical Sciences, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, Guangdong 510006, China. Electronic address: wangshengqi@gzucm.edu.cn.
  • 4 State Key Laboratory of Traditional Chinese Medicine Syndrome, Chinese Medicine Guangdong Laboratory, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510006, China; The Research Center for Integrative Medicine, School of Basic Medical Sciences, Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China. Electronic address: ellen0000@gzucm.edu.cn.
  • 5 State Key Laboratory of Traditional Chinese Medicine Syndrome, Chinese Medicine Guangdong Laboratory, The Second Affiliated Hospital of Guangzhou University of Chinese Medicine, Guangzhou 510006, China; Breast Disease Specialist Hospital of Guangdong Provincial Hospital of Chinese Medicine, the Second Clinical College of Guangzhou University of Chinese Medicine, Guangzhou, Guangdong 510006, China; Guangdong Provincial Key Laboratory of Clinical Research on Traditional Chinese Medicine Syndrome, Guangdong Provincial Academy of Chinese Medical Sciences, Guangdong Provincial Hospital of Chinese Medicine, Guangzhou, Guangdong 510006, China. Electronic address: wangzhiyu@gzucm.edu.cn.
PMID: 41072283 DOI: 10.1016/j.phymed.2025.157358
Abstract

Background: Tumor-associated macrophages (TAMs) play a pivotal role in promoting neoangiogenesis and have emerged as a promising therapeutic target in Cancer treatment. Aiduqing formula (ADQ) is a traditional Chinese medicine (TCM) formula that has demonstrated reliable clinical efficacy in breast Cancer treatment.

Purpose: This study aimed to explore the potential properties, underlying mechanisms, and bioactive constituents of ADQ in modulating breast Cancer neoangiogenesis.

Methods: A membrane-capture strategy utilizing TAM membrane-coated magnetic nanoparticles, combined with LC-MS profiling, was employed to selectively screen TAM-binding constituents from ADQ. TAMs were co-cultured with endothelial cells in vitro, and co-implanted orthotopically with breast Cancer cells in vivo to simulate their coexistence. Multiple Molecular Biology techniques, zebrafish xenotransplantation and murine xenograft models of breast Cancer were utilized to assess neoangiogenesis.

Results: ADQ administration dose-dependently reduced both microvessel density and TAM infiltration in mammary tumors. Furthermore, TAM-membrane-capture/LC-MS screening and TAM/CXCL1 ELISA assays identified liquiritin as the principal bioactive constituent of ADQ responsible for suppressing TAM/CXCL1. Moreover, liquiritin at non-toxic concentrations exerted no discernible cytotoxic effects on endothelial cell viability or function. However, it significantly and dose-dependently inhibited TAM/CXCL1-induced endothelial cell proliferation, migration, invasion, and tube formation in vitro, whereas CXCL1 overexpression in TAMs partially abrogated this effect. Mechanistically, liquiritin accumulated preferentially within TAM lysosomes, enhanced CXCL1 and lysosome co-localization, and ultimately accelerated lysosomal degradation of CXCL1 by upregulating cathepsin-K (CTSK) expression. More importantly, liquiritin significantly suppressed TAM/CXCL1-driven neoangiogenesis in the zebrafish xenotransplantation model with high-safety. Additionally, liquiritin effectively inhibited TAM/CXCL1-induced neoangiogenesis and growth of mouse breast Cancer xenografts in vivo.

Conclusion: This pioneering study not only sheds light on TAM/CXCL1 as a novel therapeutic target for suppressing breast Cancer neoangiogenesis, but also identifies liquiritin as a potent and low-toxicity anti-angiogenic phytochemical for breast Cancer treatment functioning by enhancing CTSK-mediated lysosomal degradation of CXCL1 in TAMs.

Keywords

Aiduqing formula; Breast cancer neoangiogenesis; CXCL1; Liquiritin; Tumor-associated macrophage.

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