Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice

Abstract Background The therapy for breast cancer bone metastasis (BCBM) still needs profound investigation. Focusing on the formation of bone pre-metastasis niche (BPMN) is a crucial tache for BCBM treatment. Wenshen Zhuanggu Formula (WSZG) is a typical traditional Chinese medicine (TCM) prescripti...

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Main Authors: Qionglian Huang, Hanjuan Ning, Jue Wang, Weiwei Liu, Sheng Liu, Jianyi Wang, Xianghui Han
Format: Article
Language:English
Published: BMC 2025-06-01
Series:Chinese Medicine
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Online Access:https://doi.org/10.1186/s13020-025-01136-8
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author Qionglian Huang
Hanjuan Ning
Jue Wang
Weiwei Liu
Sheng Liu
Jianyi Wang
Xianghui Han
author_facet Qionglian Huang
Hanjuan Ning
Jue Wang
Weiwei Liu
Sheng Liu
Jianyi Wang
Xianghui Han
author_sort Qionglian Huang
collection DOAJ
description Abstract Background The therapy for breast cancer bone metastasis (BCBM) still needs profound investigation. Focusing on the formation of bone pre-metastasis niche (BPMN) is a crucial tache for BCBM treatment. Wenshen Zhuanggu Formula (WSZG) is a typical traditional Chinese medicine (TCM) prescription with ability of clinically palliating bone pain and ameliorating the quality of life of BCBM patients. This study aims to elucidate the inhibitory effect and underlying mechanism of WSZG on BPMN formation induced by tumor-derived exosomes. Methods Bone precursor cells were exposed to exosomes derived from MDA-MB-231BO cells (BO-exo, 50 μg/mL) and treated by WSZG (10 or 20 μg/mL) in vitro. A primary breast cancer mouse model with propensity of BPMN was established by pre-educating BO-exo and then subcutaneously injecting MDA-MB-231BO cells into mammary fat pads. After the modeled mice were orally administrated WSZG (6.5 or 13 g crude herb kg−1 d−1) for 28 days, bone architecture and bone parameters of tibia were analyzed using micro-CT. The absorbed exosomes by bone precursor cells were investigated using confocal microscopy. Osteoclastic and osteoblastic differentiation were examined by TRAP, ALP and qRT-PCR. The levels of protein related to bone metabolism and extracellular matrix (ECM) were detected by ELISA and IHC methods. Results The results showed that BO-exo elicited the vicious bone microenvironment to form BPMN in bone-tropic primary BC mice. WSZG encouragingly targeted BO-exo to hinder osteoclastic differentiation (ACP5, c-Fos, Ctsk, MMP9, NFATc1), improve focal bone lesions (BMD, Tb.N, Tb.Th, Tb.Sp, Conn-Dens. and BV/TV), and downregulate the expression levels of bone metabolism markers (ICTP, BALP, TRACP-5b) and bone ECM proteins (bone sialoprotein, elastin, fibronectin, osteopontin, collagen I and vitronectin). Conclusions This study denotes the enhanced effects of BO-exo and highly inhibitive possibility by WSZG treatment on BPMN formation, shaping bone metastatic landscape of BC and informing the treatment of early-stage BCBM in terms of TCM.
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spelling doaj-art-44400f49daf1419c8494f18359ec5dfe2025-08-20T02:37:35ZengBMCChinese Medicine1749-85462025-06-0120111910.1186/s13020-025-01136-8Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer miceQionglian Huang0Hanjuan Ning1Jue Wang2Weiwei Liu3Sheng Liu4Jianyi Wang5Xianghui Han6Institute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese MedicineInstitute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese MedicineInstitute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese MedicineInstitute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese MedicineInstitute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese MedicineDepartment of Liver Disease, Shanghai Yueyang Integrated Traditional Chinese Medicine and Western Medicine Hospital Affiliated to Shanghai University of Traditional Chinese MedicineInstitute of Chinese Traditional Surgery, Longhua Hospital, Shanghai University of Traditional Chinese MedicineAbstract Background The therapy for breast cancer bone metastasis (BCBM) still needs profound investigation. Focusing on the formation of bone pre-metastasis niche (BPMN) is a crucial tache for BCBM treatment. Wenshen Zhuanggu Formula (WSZG) is a typical traditional Chinese medicine (TCM) prescription with ability of clinically palliating bone pain and ameliorating the quality of life of BCBM patients. This study aims to elucidate the inhibitory effect and underlying mechanism of WSZG on BPMN formation induced by tumor-derived exosomes. Methods Bone precursor cells were exposed to exosomes derived from MDA-MB-231BO cells (BO-exo, 50 μg/mL) and treated by WSZG (10 or 20 μg/mL) in vitro. A primary breast cancer mouse model with propensity of BPMN was established by pre-educating BO-exo and then subcutaneously injecting MDA-MB-231BO cells into mammary fat pads. After the modeled mice were orally administrated WSZG (6.5 or 13 g crude herb kg−1 d−1) for 28 days, bone architecture and bone parameters of tibia were analyzed using micro-CT. The absorbed exosomes by bone precursor cells were investigated using confocal microscopy. Osteoclastic and osteoblastic differentiation were examined by TRAP, ALP and qRT-PCR. The levels of protein related to bone metabolism and extracellular matrix (ECM) were detected by ELISA and IHC methods. Results The results showed that BO-exo elicited the vicious bone microenvironment to form BPMN in bone-tropic primary BC mice. WSZG encouragingly targeted BO-exo to hinder osteoclastic differentiation (ACP5, c-Fos, Ctsk, MMP9, NFATc1), improve focal bone lesions (BMD, Tb.N, Tb.Th, Tb.Sp, Conn-Dens. and BV/TV), and downregulate the expression levels of bone metabolism markers (ICTP, BALP, TRACP-5b) and bone ECM proteins (bone sialoprotein, elastin, fibronectin, osteopontin, collagen I and vitronectin). Conclusions This study denotes the enhanced effects of BO-exo and highly inhibitive possibility by WSZG treatment on BPMN formation, shaping bone metastatic landscape of BC and informing the treatment of early-stage BCBM in terms of TCM.https://doi.org/10.1186/s13020-025-01136-8Breast cancerBone pre-metastasis nicheExosomeWenshen Zhuanggu formulaBone microenvironment
spellingShingle Qionglian Huang
Hanjuan Ning
Jue Wang
Weiwei Liu
Sheng Liu
Jianyi Wang
Xianghui Han
Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice
Chinese Medicine
Breast cancer
Bone pre-metastasis niche
Exosome
Wenshen Zhuanggu formula
Bone microenvironment
title Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice
title_full Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice
title_fullStr Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice
title_full_unstemmed Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice
title_short Wenshen Zhuanggu formula inhibits tumor-exosomes induced bone pre-metastasis niche formation in primary breast cancer mice
title_sort wenshen zhuanggu formula inhibits tumor exosomes induced bone pre metastasis niche formation in primary breast cancer mice
topic Breast cancer
Bone pre-metastasis niche
Exosome
Wenshen Zhuanggu formula
Bone microenvironment
url https://doi.org/10.1186/s13020-025-01136-8
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