CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism

Abstract Background Fibrogenesis is a common pathological feature of endometriotic lesions and contributes to the development of endometriosis-associated chronic pelvic pain and infertility. TGF-β is a critical factor in the induction of fibrogenesis; however, the underlying regulatory mechanisms of...

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Main Authors: Mian Liu, Yi Gong, Hong Cai, Rui Hua, Hong Li, Zhe Wang, Yao Zhou, Yanlin Ma, Song Quan
Format: Article
Language:English
Published: BMC 2025-07-01
Series:Journal of Translational Medicine
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Online Access:https://doi.org/10.1186/s12967-025-06804-9
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author Mian Liu
Yi Gong
Hong Cai
Rui Hua
Hong Li
Zhe Wang
Yao Zhou
Yanlin Ma
Song Quan
author_facet Mian Liu
Yi Gong
Hong Cai
Rui Hua
Hong Li
Zhe Wang
Yao Zhou
Yanlin Ma
Song Quan
author_sort Mian Liu
collection DOAJ
description Abstract Background Fibrogenesis is a common pathological feature of endometriotic lesions and contributes to the development of endometriosis-associated chronic pelvic pain and infertility. TGF-β is a critical factor in the induction of fibrogenesis; however, the underlying regulatory mechanisms of TGF-β-induced fibrosis in endometriosis remain unclear. In this study, we investigated the effects and mechanisms of CCN5 in regulating the progression of TGF-β-induced fibrosis in endometriosis. Methods We investigated the role of CCN5 in TGF-β-induced proliferation and pro-fibrotic responses in primary HESCs through CCN5 overexpression and knockdown techniques. We evaluated the impact of CCN5 modulation on the activation of the TGF-β/Smad and Wnt/β-catenin signaling pathways in primary HESCs subjected to TGF-β stimulation. To elucidate the role of Smad3 in CCN5 mediating TGF-β-induced pro-fibrotic response and the activation of TGF-β/Smad and Wnt/β-catenin signaling pathways, we employed SIS3, a specific inhibitor of Smad3. Additionally, we assessed the interaction between CCN5 and Smad3 in primary HESCs. Results The expression of CCN5 was significantly elevated at both the mRNA and protein levels in patients with endometriosis compared to healthy controls. Overexpression of CCN5 through transfection with LV-CCN5 notably attenuated TGF-β-induced proliferation and pro-fibrotic responses, whereas CCN5 knockdown exhibited the opposite effects in primary HESCs. Additionally, we observed the Wnt/β-catenin signaling pathway, a classical TGF-β-associated pro-fibrotic pathway, was significantly activated in primary HESCs under TGF-β stimulation. CCN5 overexpression inhibited the increased activity of both TGF-β/Smad and Wnt/β-catenin signaling pathways induced by TGF-β, while knockdown of CCN5 significantly enhanced TGF-β-induced activation of these pathways, an effect that was partially mitigated by the TGF-β inhibitor pirfenidone. An in vitro study demonstrated that SIS3, a specific Smad3 inhibitor, blocked the effects of CCN5 knockdown on TGF-β-induced proliferation and pro-fibrotic responses in endometriosis. Furthermore, we established that CCN5 directly interacts with Smad3 in cytoplasm, inhibiting Smad3’s translocation into the nucleus and the subsequent activation of downstream target genes associated with TGF-β signaling pathways. Conclusions CCN5 serves as an crucial negative regulator of fibrosis progression in endometriosis and represents a potential therapeutic target for endometrial fibrosis.
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spelling doaj-art-cf2fa3ecaa49428388529db222c58e3c2025-08-20T03:43:02ZengBMCJournal of Translational Medicine1479-58762025-07-0123111810.1186/s12967-025-06804-9CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanismMian Liu0Yi Gong1Hong Cai2Rui Hua3Hong Li4Zhe Wang5Yao Zhou6Yanlin Ma7Song Quan8Center for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityHenan Provincial Key Laboratory of Genetic Diseases and Functional Genomics, Medical Genetic Institute of Henan Province, Henan Provincial People’s Hospital, Henan Provincial People’s Clinical Medical School of Zhengzhou UniversityDepartment of Obstetrics and Gynecology, Shenzhen Hospital of Southern Medical UniversityCenter for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityCenter for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityCenter for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityCenter for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityHainan Provincial Key Laboratory for Human Reproductive Medicine and Genetic Research, Hainan Provincial Clinical Research Center for Thalassemia, Key Laboratory of Reproductive Health Diseases Research and Translation (Hainan Medical University), Ministry of Education, Department of Reproductive Medicine, The First Affiliated Hospital of Hainan Medical University, Hainan Medical UniversityCenter for Reproductive Medicine, Department of Obstetrics and Gynecology, Nanfang Hospital, Southern Medical UniversityAbstract Background Fibrogenesis is a common pathological feature of endometriotic lesions and contributes to the development of endometriosis-associated chronic pelvic pain and infertility. TGF-β is a critical factor in the induction of fibrogenesis; however, the underlying regulatory mechanisms of TGF-β-induced fibrosis in endometriosis remain unclear. In this study, we investigated the effects and mechanisms of CCN5 in regulating the progression of TGF-β-induced fibrosis in endometriosis. Methods We investigated the role of CCN5 in TGF-β-induced proliferation and pro-fibrotic responses in primary HESCs through CCN5 overexpression and knockdown techniques. We evaluated the impact of CCN5 modulation on the activation of the TGF-β/Smad and Wnt/β-catenin signaling pathways in primary HESCs subjected to TGF-β stimulation. To elucidate the role of Smad3 in CCN5 mediating TGF-β-induced pro-fibrotic response and the activation of TGF-β/Smad and Wnt/β-catenin signaling pathways, we employed SIS3, a specific inhibitor of Smad3. Additionally, we assessed the interaction between CCN5 and Smad3 in primary HESCs. Results The expression of CCN5 was significantly elevated at both the mRNA and protein levels in patients with endometriosis compared to healthy controls. Overexpression of CCN5 through transfection with LV-CCN5 notably attenuated TGF-β-induced proliferation and pro-fibrotic responses, whereas CCN5 knockdown exhibited the opposite effects in primary HESCs. Additionally, we observed the Wnt/β-catenin signaling pathway, a classical TGF-β-associated pro-fibrotic pathway, was significantly activated in primary HESCs under TGF-β stimulation. CCN5 overexpression inhibited the increased activity of both TGF-β/Smad and Wnt/β-catenin signaling pathways induced by TGF-β, while knockdown of CCN5 significantly enhanced TGF-β-induced activation of these pathways, an effect that was partially mitigated by the TGF-β inhibitor pirfenidone. An in vitro study demonstrated that SIS3, a specific Smad3 inhibitor, blocked the effects of CCN5 knockdown on TGF-β-induced proliferation and pro-fibrotic responses in endometriosis. Furthermore, we established that CCN5 directly interacts with Smad3 in cytoplasm, inhibiting Smad3’s translocation into the nucleus and the subsequent activation of downstream target genes associated with TGF-β signaling pathways. Conclusions CCN5 serves as an crucial negative regulator of fibrosis progression in endometriosis and represents a potential therapeutic target for endometrial fibrosis.https://doi.org/10.1186/s12967-025-06804-9EndometriosisCCN5TGF-βFibrosisWnt/β-cateninSmad
spellingShingle Mian Liu
Yi Gong
Hong Cai
Rui Hua
Hong Li
Zhe Wang
Yao Zhou
Yanlin Ma
Song Quan
CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism
Journal of Translational Medicine
Endometriosis
CCN5
TGF-β
Fibrosis
Wnt/β-catenin
Smad
title CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism
title_full CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism
title_fullStr CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism
title_full_unstemmed CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism
title_short CCN5 negatively regulates TGF-β-induced endometriosis associated fibrosis through Wnt/β-catenin signaling via Smad3-dependent mechanism
title_sort ccn5 negatively regulates tgf β induced endometriosis associated fibrosis through wnt β catenin signaling via smad3 dependent mechanism
topic Endometriosis
CCN5
TGF-β
Fibrosis
Wnt/β-catenin
Smad
url https://doi.org/10.1186/s12967-025-06804-9
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