si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway
Abstract Background Intrauterine adhesions (IUAs) are manifestations of endometrial fibrosis characterized by inflammation and fibrinogen aggregation in the extracellular matrix (ECM). The available therapeutic interventions for IUA are insufficiently effective in the clinical setting for postoperat...
Main Authors: | , , , , , , , , |
---|---|
Format: | Article |
Language: | English |
Published: |
BMC
2020-11-01
|
Series: | Stem Cell Research & Therapy |
Subjects: | |
Online Access: | http://link.springer.com/article/10.1186/s13287-020-01990-3 |
id |
doaj-88aba6a5bb38493585cf2b1d19ba9e7a |
---|---|
record_format |
Article |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Limin Liu Guobin Chen Taoliang Chen Wenjuan Shi Haiyan Hu Kaijing Song Ruichun Huang Huihua Cai Yuanli He |
spellingShingle |
Limin Liu Guobin Chen Taoliang Chen Wenjuan Shi Haiyan Hu Kaijing Song Ruichun Huang Huihua Cai Yuanli He si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway Stem Cell Research & Therapy IUA TGF-β1 FOXF2 SNHG5 ECM Fibrosis |
author_facet |
Limin Liu Guobin Chen Taoliang Chen Wenjuan Shi Haiyan Hu Kaijing Song Ruichun Huang Huihua Cai Yuanli He |
author_sort |
Limin Liu |
title |
si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway |
title_short |
si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway |
title_full |
si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway |
title_fullStr |
si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway |
title_full_unstemmed |
si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathway |
title_sort |
si-snhg5-foxf2 inhibits tgf-β1-induced fibrosis in human primary endometrial stromal cells by the wnt/β-catenin signalling pathway |
publisher |
BMC |
series |
Stem Cell Research & Therapy |
issn |
1757-6512 |
publishDate |
2020-11-01 |
description |
Abstract Background Intrauterine adhesions (IUAs) are manifestations of endometrial fibrosis characterized by inflammation and fibrinogen aggregation in the extracellular matrix (ECM). The available therapeutic interventions for IUA are insufficiently effective in the clinical setting for postoperative adhesion recurrence and infertility problems. In this study, we investigated whether si-SNHG5-FOXF2 can serve as a molecular mechanism for the inhibition of IUA fibrosis ex vivo. Methods FOXF2, TGF-β1 and collagen expression levels were measured by microarray sequencing analysis in three normal endometrium groups and six IUA patients. We induced primary human endometrial stromal cells (HESCs) into myofibroblasts (MFs) to develop an IUA cell model with various concentrations of TGF-β1 at various times. Downstream target genes of FOXF2 were screened by chromatin immunoprecipitation combined with whole-genome high-throughput sequencing (ChIP-seq). We investigated ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related proteins in primary HESCs with FOXF2 downregulation by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting (WB), immunohistochemistry (IHC), flow cytometry, ethylenediurea (EdU) and CCK8 assays. We identified long noncoding RNAs (lncRNA) SNHG5 as the upstream regulatory gene of FOXF2 through RNA immunoprecipitation (RIP), RNA pulldown and fluorescence in situ hybridization (FISH). Finally, we examined FOXF2 expression, ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related proteins in primary HESCs upon FOXF2 downregulation. Results FOXF2 was highly expressed in the endometrium of patients with IUA. Treatment of primary HESCs with 10 ng/ml TGF-β1 for 72 h was found to be most effective for developing an IUA cell model. FOXF2 regulated multiple downstream target genes, including collagen, vimentin (VIM) and cyclin D2/DK4, by ChIP-seq and ChIP-PCR. FOXF2 downregulation inhibited TGF-β1-mediated primary HESC fibrosis, including ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related protein expression. We identified lncRNA SNHG5 as an upstream gene that directly regulates FOXF2 by RIP-seq, qRT-PCR, WB and FISH. SNHG5 downregulation suppressed FOXF2 expression in the IUA cell model, resulting in synergistic repression of the Wnt/β-catenin pathway, thereby altering TGF-β1-mediated ECM aggregation in endometrial stromal cells ex vivo. Conclusions Regulation of the Wnt/β-catenin signalling pathway and ECM formation by si-SNHG5-FOXF2 effectively inhibited the profibrotic effect of TGF-β1 on primary HESCs. This finding can provide a molecular basis for antagonizing TGF-β1-mediated fibrosis in primary HESCs. |
topic |
IUA TGF-β1 FOXF2 SNHG5 ECM Fibrosis |
url |
http://link.springer.com/article/10.1186/s13287-020-01990-3 |
work_keys_str_mv |
AT liminliu sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT guobinchen sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT taoliangchen sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT wenjuanshi sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT haiyanhu sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT kaijingsong sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT ruichunhuang sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT huihuacai sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway AT yuanlihe sisnhg5foxf2inhibitstgfb1inducedfibrosisinhumanprimaryendometrialstromalcellsbythewntbcateninsignallingpathway |
_version_ |
1724422848650412032 |
spelling |
doaj-88aba6a5bb38493585cf2b1d19ba9e7a2020-11-25T04:09:12ZengBMCStem Cell Research & Therapy1757-65122020-11-0111111710.1186/s13287-020-01990-3si-SNHG5-FOXF2 inhibits TGF-β1-induced fibrosis in human primary endometrial stromal cells by the Wnt/β-catenin signalling pathwayLimin Liu0Guobin Chen1Taoliang Chen2Wenjuan Shi3Haiyan Hu4Kaijing Song5Ruichun Huang6Huihua Cai7Yuanli He8Department of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical UniversityDepartment of Obstetrics and Gynecology, Affiliated Shenzhen Maternity & Child Healthcare Hospital, Southern Medical UniversityThe National Key Clinical Specialty, The Engineering Technology Research Center of Education Ministry of China, Guangdong Provincial Key Laboratory on Brain Function Repair and Regeneration, Department of Neurosurgery, Zhujiang Hospital, Southern Medical UniversityDepartment of Obstetrics and Gynecology, Affiliated Shenzhen Maternity & Child Healthcare Hospital, Southern Medical UniversityDepartment of Obstetrics and Gynecology, Affiliated Shenzhen Maternity & Child Healthcare Hospital, Southern Medical UniversityDepartment of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical UniversityDepartment of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical UniversityDepartment of Obstetrics and Gynecology, Guangdong Provincial People’s Hospital, Guangdong Academy of Medical SciencesDepartment of Obstetrics and Gynecology, Zhujiang Hospital, Southern Medical UniversityAbstract Background Intrauterine adhesions (IUAs) are manifestations of endometrial fibrosis characterized by inflammation and fibrinogen aggregation in the extracellular matrix (ECM). The available therapeutic interventions for IUA are insufficiently effective in the clinical setting for postoperative adhesion recurrence and infertility problems. In this study, we investigated whether si-SNHG5-FOXF2 can serve as a molecular mechanism for the inhibition of IUA fibrosis ex vivo. Methods FOXF2, TGF-β1 and collagen expression levels were measured by microarray sequencing analysis in three normal endometrium groups and six IUA patients. We induced primary human endometrial stromal cells (HESCs) into myofibroblasts (MFs) to develop an IUA cell model with various concentrations of TGF-β1 at various times. Downstream target genes of FOXF2 were screened by chromatin immunoprecipitation combined with whole-genome high-throughput sequencing (ChIP-seq). We investigated ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related proteins in primary HESCs with FOXF2 downregulation by quantitative reverse transcription-polymerase chain reaction (qRT-PCR), western blotting (WB), immunohistochemistry (IHC), flow cytometry, ethylenediurea (EdU) and CCK8 assays. We identified long noncoding RNAs (lncRNA) SNHG5 as the upstream regulatory gene of FOXF2 through RNA immunoprecipitation (RIP), RNA pulldown and fluorescence in situ hybridization (FISH). Finally, we examined FOXF2 expression, ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related proteins in primary HESCs upon FOXF2 downregulation. Results FOXF2 was highly expressed in the endometrium of patients with IUA. Treatment of primary HESCs with 10 ng/ml TGF-β1 for 72 h was found to be most effective for developing an IUA cell model. FOXF2 regulated multiple downstream target genes, including collagen, vimentin (VIM) and cyclin D2/DK4, by ChIP-seq and ChIP-PCR. FOXF2 downregulation inhibited TGF-β1-mediated primary HESC fibrosis, including ECM formation, cell proliferation and Wnt/β-catenin signalling pathway-related protein expression. We identified lncRNA SNHG5 as an upstream gene that directly regulates FOXF2 by RIP-seq, qRT-PCR, WB and FISH. SNHG5 downregulation suppressed FOXF2 expression in the IUA cell model, resulting in synergistic repression of the Wnt/β-catenin pathway, thereby altering TGF-β1-mediated ECM aggregation in endometrial stromal cells ex vivo. Conclusions Regulation of the Wnt/β-catenin signalling pathway and ECM formation by si-SNHG5-FOXF2 effectively inhibited the profibrotic effect of TGF-β1 on primary HESCs. This finding can provide a molecular basis for antagonizing TGF-β1-mediated fibrosis in primary HESCs.http://link.springer.com/article/10.1186/s13287-020-01990-3IUATGF-β1FOXF2SNHG5ECMFibrosis |