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©The Author(s) 2024. Published by Baishideng Publishing Group Inc. All rights reserved.
Calculus bovis inhibits M2 tumor-associated macrophage polarization via Wnt/β-catenin pathway modulation to suppress liver cancer
Zhen Huang, Fan-Ying Meng, Lin-Zhu Lu, Qian-Qian Guo, Chang-Jun Lv, Nian-Hua Tan, Zhe Deng, Jun-Yi Chen, Zi-Shu Zhang, Bo Zou, Hong-Ping Long, Qing Zhou, Sha Tian, Si Mei, Xue-Fei Tian
Zhen Huang, Lin-Zhu Lu, Qian-Qian Guo, Chang-Jun Lv, Nian-Hua Tan, Zhe Deng, Jun-Yi Chen, Sha Tian, Xue-Fei Tian, College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Zhen Huang, Lin-Zhu Lu, Qian-Qian Guo, Chang-Jun Lv, Nian-Hua Tan, Jun-Yi Chen, Si Mei, Xue-Fei Tian, Hunan Key Laboratory of Translational Research in Formulas and Zheng of Traditional Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Zhen Huang, Lin-Zhu Lu, Qian-Qian Guo, Chang-Jun Lv, Nian-Hua Tan, Jun-Yi Chen, Si Mei, Xue-Fei Tian, Key Laboratory of Traditional Chinese Medicine for Mechanism of Tumor Prevention and Treatment, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Fan-Ying Meng, Zi-Shu Zhang, Bo Zou, The First Clinical College of Traditional Chinese Medicine, Hunan University of Traditional Chinese Medicine, Changsha 410007, Hunan Province, China
Nian-Hua Tan, Department of Hepatology, Hunan University of Chinese Medicine, Changsha 410007, Hunan Province, China
Hong-Ping Long, Qing Zhou, The First Hospital of Hunan University of Chinese Medicine, Hunan University of Chinese Medicine, Changsha 410007, Hunan Province, China
Si Mei, Faculty of Medicine, Hunan University of Chinese Medicine, Changsha 410208, Hunan Province, China
Co-first authors: Zhen Huang and Fan-Ying Meng.
Co-corresponding authors: Si Mei and Xue-Fei Tian.
Author contributions: Huang Z, Meng FY, Mei S, and Tian XF designed the study; Huang Z, Meng FY, Lu LZ, Guo QQ, Deng Z, Zou B, and Long HP performed the experiments, and acquired and analyzed the data; Huang Z, Meng FY, Lu LZ, and Guo QQ prepared the figures and tables; Huang Z, Meng FY, Lu LZ, Guo QQ, Lv CJ, Tan NH, Deng Z, Chen JY, Zhang ZS, Zhou Q, Mei S, and Tian XF reviewed and edited the manuscript. All authors reviewed and approved the final version of the article. Huang Z and Meng FY are designated as co-first authors due to their nearly equal contributions across various aspects of the project, including study design, data collection and analysis, and manuscript writing. Their dedication and efforts in these crucial stages highlight their high level of collaboration and professional competence, which has ensured the smooth progress and high quality of the research. The comparable workload and impact of both authors justify their co-first author designation, reflecting their significant contributions fairly. Additionally, the research team comprises members with diverse expertise and skills across multiple disciplines. This diversity forms a solid foundation for the study, ensuring its comprehensiveness and reliability. The designation of co-first authors not only exemplifies the collaborative spirit and professional standards within the team but also enhances the scientific rigor and integrity of the paper. The reasons for designating Tian XF and Mei S as co-corresponding authors are threefold. First, both authors have made equally important contributions to the research work and have invested equal amounts of effort in writing the paper and designing the experiments. To be fair, they are listed as co-corresponding authors. Second, both authors have made significant contributions in their respective fields and have assumed different responsibilities in the paper. Their joint communication better reflects the nature of interdisciplinary cooperation. Third, having two corresponding authors provides a wider range of contact channels for readers to communicate and discuss with the research team, improving the dissemination and influence of the research work.
Supported by National Natural Science Foundation of China, No. 82074450; Education Department of Hunan Province, No. 21A0243, No. 21B0374, No. 22B0397, and No. 22B0392; Research Project of "Academician Liu Liang Workstation" of Hunan University of Traditional Chinese Medicine, No. 21YS003; Hunan Administration of Traditional Chinese Medicine, No. B2023001 and No. B2023009; and Hunan Provincial Natural Science Foundation of China, No. 2023JJ40481.
Institutional animal care and use committee statement: All animal experiments were conducted according to a protocol approved by the Ethical Review Committee of Experimental Animal Welfare at Slacker Jingda Laboratory, Changsha, Hunan (No. IACUC-SJA2022105).
Conflict-of-interest statement: The authors declare that they have no competing interests to disclose.
Data sharing statement: Dataset is available from the corresponding author at
003640@hnucm.edu.cn. Participants gave informed consent for data sharing.
ARRIVE guidelines statement: The authors have read the ARRIVE guidelines, and the manuscript was prepared and revised according to the ARRIVE guidelines.
Open-Access: This article is an open-access article that was selected by an in-house editor and fully peer-reviewed by external reviewers. It is distributed in accordance with the Creative Commons Attribution NonCommercial (CC BY-NC 4.0) license, which permits others to distribute, remix, adapt, build upon this work non-commercially, and license their derivative works on different terms, provided the original work is properly cited and the use is non-commercial. See:
Https://creativecommons.org/Licenses/by-nc/4.0/ Corresponding author: Xue-Fei Tian, College of Integrated Chinese and Western Medicine, Hunan University of Chinese Medicine, No. 300 Xueshi Road, Changsha 410208, Hunan Province, China.
003640@hnucm.edu.cn
Received: April 9, 2024
Revised: June 5, 2024
Accepted: July 5, 2024
Published online: August 7, 2024
Processing time: 110 Days and 23.5 Hours
BACKGROUND
Calculus bovis (CB), used in traditional Chinese medicine, exhibits anti-tumor effects in various cancer models. It also constitutes an integral component of a compound formulation known as Pien Tze Huang, which is indicated for the treatment of liver cancer. However, its impact on the liver cancer tumor microenvironment, particularly on tumor-associated macrophages (TAMs), is not well understood.
AIM
To elucidate the anti-liver cancer effect of CB by inhibiting M2-TAM polarization via Wnt/β-catenin pathway modulation.
METHODS
This study identified the active components of CB using UPLC-Q-TOF-MS, evaluated its anti-neoplastic effects in a nude mouse model, and elucidated the underlying mechanisms via network pharmacology, transcriptomics, and molecular docking. In vitro assays were used to investigate the effects of CB-containing serum on HepG2 cells and M2-TAMs, and Wnt pathway modulation was validated by real-time reverse transcriptase-polymerase chain reaction and Western blot analysis.
RESULTS
This study identified 22 active components in CB, 11 of which were detected in the bloodstream. Preclinical investigations have demonstrated the ability of CB to effectively inhibit liver tumor growth. An integrated approach employing network pharmacology, transcriptomics, and molecular docking implicated the Wnt signaling pathway as a target of the antineoplastic activity of CB by suppressing M2-TAM polarization. In vitro and in vivo experiments further confirmed that CB significantly hinders M2-TAM polarization and suppresses Wnt/β-catenin pathway activation. The inhibitory effect of CB on M2-TAMs was reversed when treated with the Wnt agonist SKL2001, confirming its pathway specificity.
CONCLUSION
This study demonstrated that CB mediates inhibition of M2-TAM polarization through the Wnt/β-catenin pathway, contributing to the suppression of liver cancer growth.
Core Tip: Calculus bovis (CB), a valuable herb in traditional Chinese medicine, has shown definite anti-liver cancer effects in vivo. By analyzing the composition of CB and using network pharmacology for target prediction, we found that CB exhibits anti-liver cancer effects by affecting immune-related pathways in the tumor microenvironment. Through transcriptome sequencing, we further showed that regulation of the M2-type polarization of tumor-associated macrophages (TAMs) is responsible for the effects of CB. In vitro studies showed that modulating the Wnt/β-catenin pathway is a crucial mechanism by which CB regulates M2 polarization of TAMs. This study provides evidence for the development of anti-liver cancer drugs.
