Skip to main content
Key Specialties
Cardiology Diabetology Endocrinology Gastroenterology Geriatrics and Gerontology Gynecology and Obstetrics Hematology Infectious Disease Internal Medicine Nephrology Neurology Oncology Pediatrics Respiratory Medicine Rheumatology Surgery
Congress Coverage
2024 ACC Congress 2023 ESMO Congress 2023 EASD Congress 2023 ERS Congress 2023 ESC Congress
Clinical Collections
Advances in Alzheimer’s

At a glance: The ONWARDS insulin icodec trials

A round-up of the ONWARDS phase 3 clinical trials evaluating the novel weekly insulin icodec in people with diabetes.
ADVANCED SEARCH
Log in
Top
BMC Urology
Published in: BMC Urology 1/2020

01-12-2020 | Multiple Sclerosis | Research article

MicroRNA-219c-5p regulates bladder fibrosis by targeting FN1

Authors: Bowen Liu, Yafei Ding, Peng Li, Tao Wang, Siyuan He, Zhankui Jia, Jinjian Yang

Published in: BMC Urology | Issue 1/2020

Login to get access

Abstract

Background

We found that the bladders of multiple sclerosis mice were significantly fibrotic. This study aimed to investigate the relationship between fibronectin 1 (FN1) and bladder fibrosis, as well as the microRNAs involved in FN1 regulation.

Methods

The degree of bladder smooth muscle fibrosis was observed by immunohistochemistry. In addition, we used quantitative real-time polymerase chain reaction (RT-qPCR) and Western blotting to determine FN1 expression in bladders with different grades of fibrosis. Bioinformatics analysis revealed that miR-199a-3p, miR-219c-5p and miR-3572-3p could inhibit FN1 synthesis. Therefore, miR-199a-3p, miR-219c-5p and miR-3572-3p were overexpressed or knocked down in bladder smooth muscle cells (BSMCs), and the respective transfection and FN1 knockdown efficiencies were detected by RT-qPCR. Only miR-219c-5p overexpression and knockdown produced the expected results. A dual luciferase reporter assay was used to determine the targeting relationship between miR-219c-5p and FN1. Flow cytometry and Cell Counting Kit 8 (CCK8) experiments confirmed that miR-219c-5p reduced FN1 expression and affected the biological activity of smooth muscle cells. Agomir and anagomir of miR-219c-5p were transfected in vivo to observe the change of bladder fibrosis in mice.

Results

With increasing bladder fibrosis, FN1 expression increased, while miR-199a-3p, miR-219c-5p, and miR-3572-3p expression levels decreased. The RT-qPCR results after transfection showed that only miR-219c-5p could regulate FN1. Indeed, the dual luciferase reporter assay results indicated that miR-219c-5p targeted FN1 directly. CCK8 and cell cycle assays showed that miR-219c-5p overexpression inhibited BSMC proliferation, while miR-219c-5p knockdown promoted BSMC proliferation. An apoptosis assay showed that miR-219c-5p overexpression promoted apoptosis, while miR-219c-5p knockdown inhibited BSMC apoptosis. The agomir and anagomir transfected with miR-219c-5p in vivo found that the bladder fibrosis of the mice in the agomir group was reduced, and the anagomir group was worse.

Conclusions

Our findings indicate that FN1 up-regulation and miR-219c-5p down-regulation play an important role in the development of bladder fibrosis, and miR-219c-5p participates in bladder fibrosis by regulating FN1 expression. Thus, a novel antifibrotic function of miR-219c-5p is proposed, which may represent a potential target for the diagnosis and treatment of bladder fibrosis.
Literature
1.
Hafler DA, Slavik JM, Anderson DE, O’Connor KC, De Jager P, Baecher-Allan C. Multiple sclerosis. Immunol Rev. 2005;204:208–31.PubMedCrossRef
2.
3.
Fernandez O. Mechanisms and current treatments of urogenital dysfunction in multiple sclerosis. J Neurol. 2002;249(1):1–8.PubMedCrossRef
4.
Kalsi V, Fowler CJ. Therapy Insight: bladder dysfunction associated with multiple sclerosis. Nat Clin Pract Urol. 2005;2(10):492–501.PubMedCrossRef
5.
Litwiller SE, Frohman EM, Zimmern PE. Multiple sclerosis and the urologist. J Urol. 1999;161(3):743–57.PubMedCrossRef
6.
Al-Izki S, Pryce G, Giovannoni G, Baker D. Evaluating potential therapies for bladder dysfunction in a mouse model of multiple sclerosis with high-resolution ultrasonography. Mult Scler. 2009;15(7):795–801.PubMedCrossRef
7.
Altuntas CZ, Daneshgari F, Izgi K, Bicer F, Ozer A, Sakalar C, Grimberg KO, Sayin I, Tuohy VK. Connective tissue and its growth factor CTGF distinguish the morphometric and molecular remodeling of the bladder in a model of neurogenic bladder. Am J Physiol Renal Physiol. 2012;303(9):F1363-1369.PubMedCrossRefPubMedCentral
8.
Steinman L, Zamvil SS. Virtues and pitfalls of EAE for the development of therapies for multiple sclerosis. Trends Immunol. 2005;26(11):565–71.PubMedCrossRef
9.
Petry KG, Boullerne AI, Pousset F, Brochet B, Caille JM, Dousset V. Experimental allergic encephalomyelitis animal models for analyzing features of multiple sclerosis. Pathol Biol (Paris). 2000;48(1):47–53.
10.
Altuntas CZ, Daneshgari F, Liu G, Fabiyi A, Kavran M, Johnson JM, Gulen MF, Jaini R, Li X, Frenkl TL, et al. Bladder dysfunction in mice with experimental autoimmune encephalomyelitis. J Neuroimmunol. 2008;203(1):58–63.PubMedCrossRefPubMedCentral
11.
Anumanthan G, Tanaka ST, Adams CM, Thomas JC, Wills ML, Adams MC, Hayward SW, Matusik RJ, Bhowmick NA, Brock JW 3rd, et al. Bladder stromal loss of transforming growth factor receptor II decreases fibrosis after bladder obstruction. J Urol. 2009;182(4 Suppl):1775–80.PubMedCrossRefPubMedCentral
12.
Taylor JA, Zhu Q, Irwin B, Maghaydah Y, Tsimikas J, Pilbeam C, Leng L, Bucala R, Kuchel GA. Null mutation in macrophage migration inhibitory factor prevents muscle cell loss and fibrosis in partial bladder outlet obstruction. Am J Physiol Renal Physiol. 2006;291(6):F1343-1353.PubMedCrossRef
13.
Slama A, Khouni H, Sriha B, Brini K, Ben Sorba N, Taher Mosbah A. Bladder fibrosis caused by eosinophilic cystitis. Ann Urol (Paris). 2003;37(5):272–4.CrossRef
14.
Wang N, Duan L, Ding J, Cao Q, Qian S, Shen H, Qi J. MicroRNA-101 protects bladder of BOO from hypoxia-induced fibrosis by attenuating TGF-beta-smad2/3 signaling. IUBMB Life. 2019;71(2):235–43.PubMed
15.
Siddiqui NY, Helfand BT, Andreev VP, Kowalski JT, Bradley MS, Lai HH, Berger MB, Mueller MG, Bickhaus JA, Packiam VT, et al. Biomarkers implicated in lower urinary tract symptoms: systematic review and pathway analyses. J Urol. 2019;2019:101097JU0000000000000257.
16.
Andersson KE, Fry C, Panicker J, Rademakers K. Which molecular targets do we need to focus on to improve lower urinary tract dysfunction? ICI-RS 2017. Neurourol Urodyn. 2018;37(S4):S117-s126.PubMedCrossRef
17.
Peyronnet B, Mironska E, Chapple C, Cardozo L, Oelke M, Dmochowski R, Amarenco G, Game X, Kirby R, Van Der Aa F, et al. A comprehensive review of overactive bladder pathophysiology: on the way to tailored treatment. Eur Urol. 2019;2019:1.
18.
Sobolewski P, Murthy NS, Kohn J, El Fray M. Adsorption of fibrinogen and fibronectin on elastomeric poly(butylene succinate) copolyesters. Langmuir. 2019;2019:1.
19.
Stribos EGD, Seelen MA, van Goor H, Olinga P, Mutsaers HAM. Murine precision-cut kidney slices as an model to evaluate the role of transforming growth factor-β1 signaling in the onset of renal fibrosis. Front Physiol. 2017;8:1026.PubMedCrossRefPubMedCentral
20.
Wang W, Jia Y-J, Yang Y-L, Xue M, Zheng Z-J, Wang L, Xue Y-M. LncRNA GAS5 exacerbates renal tubular epithelial fibrosis by acting as a competing endogenous RNA of miR-96-5p. Biomed Pharmacother. 2020;121:109411.PubMedCrossRef
21.
Rupaimoole R, Slack FJ. MicroRNA therapeutics: towards a new era for the management of cancer and other diseases. Nat Rev Drug Discov. 2017;16(3):203–22.PubMedCrossRef
22.
Bruinsma IB, van Dijk M, Bridel C, van de Lisdonk T, Haverkort SQ, Runia TF, Steinman L, Hintzen RQ, Killestein J, Verbeek MM, et al. Regulator of oligodendrocyte maturation, miR-219, a potential biomarker for MS. Journal of neuroinflammation. 2017;14(1):235.PubMedCrossRefPubMedCentral
23.
Yang L, Dong C, Yang J, Yang L, Chang N, Qi C, Li L. MicroRNA-26b-5p inhibits mouse liver fibrogenesis and angiogenesis by targeting PDGF receptor-beta. Mol Ther Nucl Acids. 2019;16:206–17.CrossRef
24.
Zhang H, Sun Z, Li Y, Fan D, Jiang H. MicroRNA-200c binding to FN1 suppresses the proliferation, migration and invasion of gastric cancer cells. Biomed Pharmacother. 2017;88:285–92.PubMedCrossRef
25.
Si M, Wang Q, Li Y, Lin H, Luo D, Zhao W, Dou X, Liu J, Zhang H, Huang Y, et al. Inhibition of hyperglycolysis in mesothelial cells prevents peritoneal fibrosis. Sci Transl Med. 2019;11:495.CrossRef
26.
Xue R, Jia Z, Kong X, Pi G, Ma S, Yang J. Effects of PGE2 EP3/EP4 receptors on bladder dysfunction in mice with experimental autoimmune encephalomyelitis. Am J Physiol Renal Physiol. 2013;305(12):F1656-1662.PubMedCrossRef
27.
Lai J, Ai J, Luo D, Jin T, Liao B, Zhou L, Feng S, Jin X, Li H, Wang K. Beta-Adrenoceptor signaling regulates proliferation and contraction of human bladder smooth muscle cells under pathological hydrostatic pressure. J Cell Biochem. 2019;2019:1.
28.
Guo JH, Su C, Jiang SY, Wang F, Feng X, Wang JT. MicroRNA-1 regulates fibronectin expression in human trabecular meshwork cells under oxidative stress. Zhonghua Yan Ke Za Zhi. 2019;55(5):355–60.PubMed
29.
Ma L, Ma J, Ou HL. MicroRNA219 overexpression serves a protective role during liver fibrosis by targeting tumor growth factor beta receptor 2. Mol Med Rep. 2019;19(3):1543–50.PubMed
30.
Duan LJ, Qi J, Kong XJ, Huang T, Qian XQ, Xu D, Liang JH, Kang J. MiR-133 modulates TGF-β1-induced bladder smooth muscle cell hypertrophic and fibrotic response: implication for a role of microRNA in bladder wall remodeling caused by bladder outlet obstruction. Cell Signal. 2015;27(2):215–27.PubMedCrossRef
31.
Gunadi BNYP, Kalim AS, Santiko W, Musthofa FD, Iskandar K, Makhmudi A. Aberrant expressions of miRNA-206 target, FN1, in multifactorial Hirschsprung disease. Orphanet J Rare Dis. 2019;14(1):5.PubMedCrossRefPubMedCentral
32.
Li B, Shen W, Peng H, Li Y, Chen F, Zheng L, Xu J, Jia L. Fibronectin 1 promotes melanoma proliferation and metastasis by inhibiting apoptosis and regulating EMT. OncoTargets and therapy. 2019;12:3207–21.PubMedCrossRefPubMedCentral
33.
Wu J, Liu S, Yuan ZW, Liu JH, Li JM, Chen T, Fang KW. MicroRNA-27a suppresses detrusor fibrosis in streptozotocin-induced diabetic rats by targeting PRKAA2 through the TGF-beta1/Smad3 signaling pathway. Cell Physiol Biochem. 2018;45(4):1333–49.PubMedCrossRef
34.
Moon SJ, Bae JM, Park KS, Tagkopoulos I, Kim KJ. Compendium of skin molecular signatures identifies key pathological features associated with fibrosis in systemic sclerosis. Ann Rheum Dis. 2019;78(6):817–25.PubMedCrossRef
35.
Tao Y, Zhang H, Huang S, Pei L, Feng M, Zhao X, Ouyang Z, Yao S, Jiang R, Wei K. miR-199a-3p promotes cardiomyocyte proliferation by inhibiting Cd151 expression. Biochem Biophys Res Commun. 2019;516(1):28–36.PubMedCrossRef
Metadata
Title
MicroRNA-219c-5p regulates bladder fibrosis by targeting FN1
Authors
Bowen Liu
Yafei Ding
Peng Li
Tao Wang
Siyuan He
Zhankui Jia
Jinjian Yang
Publication date
01-12-2020
Publisher
BioMed Central
Published in
BMC Urology / Issue 1/2020
Electronic ISSN: 1471-2490
DOI
https://doi.org/10.1186/s12894-020-00765-5

Other articles of this Issue 1/2020

BMC Urology 1/2020 Go to the issue

Research article

Critical causes in severe bleeding requiring angioembolization after percutaneous nephrolithotomy

Case report

Primary pure angiosarcoma of the testis: a vanishingly rare malignancy. Case report and literature review

Research article

The effect of preoperative ureteral stenting in retrograde Intrarenal surgery: a multicenter, propensity score-matched study

Case report

Atypical focal xanthogranulomatous pyelonephritis without clinical symptoms presenting as infiltrative renal cancer: a case report and literature review

Research article

Contasure-needleless single incision slings versus transobturator slings (TOT/TVT-O) for female patients with stress urinary incontinence: a systematic review and meta-analysis

Research article

Elevated non-high-density lipoprotein cholesterol corresponds to a high risk of nephrolithiasis in children