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Published in: World Journal of Urology 2/2018

01-02-2018 | Original Article

Identification of key genes and construction of microRNA–mRNA regulatory networks in bladder smooth muscle cell response to mechanical stimuli using microarray expression profiles and bioinformatics analysis

Authors: Liao Peng, De-Yi Luo

Published in: World Journal of Urology | Issue 2/2018

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Abstract

Purpose

To identify keys genes and elucidate miRNA–mRNA regulatory networks in Bladder smooth muscle cell (BSMC) response to mechanical stimuli.

Methods

Human BSMCs, seeded on a silicone membrane, were subjected to mechanical stretch or without stretch. Microarray was used to analyze the differential expression of mRNAs and miRNAs between human BSMCs under mechanical stretch and control static control group. Differentially expressed genes(DEGs) and miRNAs (DEMs) in these two groups were identified. Subsequently, differentially expressed DEGs were conducted with functional analysis, and then PPI network was constructed. Finally, miRNA–mRNA regulatory network was visualized using Cytoscape.

Results

1639 significant DEGs and three DEMs were identified between the stretch group and control static group. The PPI network of DEGs was constructed by STRING, which was composed of 1459 nodes and 1481 edges, including 188 upregulated genes and 255 downregulated genes. Moreover, 36 genes in the PPI network were identified as hub genes in BSMCs response to mechanical stretch, e.g. CCNH, CPSF2, TSNAX, ARPC5 and PSMD3 genes. Subsequently, 39 clusters were selected from PPI network using MCODE, and it was shown that the most significant cluster consisted of 14 nodes and 91 edges. Besides, miR-503HG was the most significantly downregulated miRNA and was predicted to target five upregulated genes, including SMAD7, CCND3, WIPI2, NYNRIN and PVRL1.

Conclusions

Our data mining and integration help reveal the mechanotransduction mechanism of BSMCs’ response to mechanical stimulation and contribute to the early diagnosis of bladder outlet obstruction (BOO) as well as the improvement of pathogenesis of BOO treatment.
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Literature
1.
go back to reference Korossis S, Bolland F, Ingham E, Fisher J, Kearney J, Southgate J (2006) Review: tissue engineering of the urinary bladder: considering structure-function relationships and the role of mechanotransduction. Tissue Eng 12(4):635–644CrossRefPubMed Korossis S, Bolland F, Ingham E, Fisher J, Kearney J, Southgate J (2006) Review: tissue engineering of the urinary bladder: considering structure-function relationships and the role of mechanotransduction. Tissue Eng 12(4):635–644CrossRefPubMed
2.
go back to reference Shyu KG (2009) Cellular and molecular effects of mechanical stretch on vascular cells and cardiac myocytes. Clin Sci (Lond) 116(5):377–389CrossRef Shyu KG (2009) Cellular and molecular effects of mechanical stretch on vascular cells and cardiac myocytes. Clin Sci (Lond) 116(5):377–389CrossRef
3.
go back to reference Coplen DE, Macarak EJ, Howard PS (2003) Matrix synthesis by bladder smooth muscle cells is modulated by stretch frequency. Vitro Cell Dev Biol Anim 39(3–4):157–162CrossRef Coplen DE, Macarak EJ, Howard PS (2003) Matrix synthesis by bladder smooth muscle cells is modulated by stretch frequency. Vitro Cell Dev Biol Anim 39(3–4):157–162CrossRef
4.
go back to reference Huang H, Kamm RD, Lee RT (2004) Cell mechanics and mechanotransduction: pathways, probes, and physiology. Am J Physiol Cell Physiol 287(1):C1–C11CrossRefPubMed Huang H, Kamm RD, Lee RT (2004) Cell mechanics and mechanotransduction: pathways, probes, and physiology. Am J Physiol Cell Physiol 287(1):C1–C11CrossRefPubMed
6.
go back to reference Luo DY, Wazir R, Du C, Tian Y, Yue X, Wei TQ, Wang KJ (2015) Magnitude-dependent proliferation and contractility modulation of human bladder smooth muscle cells under physiological stretch. World J Urol 33(11):1881–1887CrossRefPubMed Luo DY, Wazir R, Du C, Tian Y, Yue X, Wei TQ, Wang KJ (2015) Magnitude-dependent proliferation and contractility modulation of human bladder smooth muscle cells under physiological stretch. World J Urol 33(11):1881–1887CrossRefPubMed
7.
go back to reference Chen S, Peng C, Wei X, Luo D, Lin Y, Yang T, Jin X, Gong L, Li H, Wang K (2017) Simulated physiological stretch increases expression of extracellular matrix proteins in human bladder smooth muscle cells via integrin α4/αv-FAK-ERK1/2 signaling pathway. World J Urol 35(8):1247–1254CrossRefPubMed Chen S, Peng C, Wei X, Luo D, Lin Y, Yang T, Jin X, Gong L, Li H, Wang K (2017) Simulated physiological stretch increases expression of extracellular matrix proteins in human bladder smooth muscle cells via integrin α4/αv-FAK-ERK1/2 signaling pathway. World J Urol 35(8):1247–1254CrossRefPubMed
8.
go back to reference Luo DY, Wazir R, Tian Y, Yue X, Wei TQ, Wang KJ (2013) Integrin αv mediates contractility whereas integrin α4 regulates proliferation of human bladder smooth muscle cells via FAK pathway under physiological stretch. J Urol 190(4):1421–1429CrossRefPubMed Luo DY, Wazir R, Tian Y, Yue X, Wei TQ, Wang KJ (2013) Integrin αv mediates contractility whereas integrin α4 regulates proliferation of human bladder smooth muscle cells via FAK pathway under physiological stretch. J Urol 190(4):1421–1429CrossRefPubMed
10.
go back to reference Boopathi E, Gomes C, Zderic SA, Malkowicz B, Chakrabarti R, Patel DP, Wein AJ, Chacko S (2014) Mechanical stretch upregulates proteins involved in Ca2+ sensitization in urinary bladder smooth muscle hypertrophy. Am J Physiol Cell Physiol 307(6):C542–C553CrossRefPubMedPubMedCentral Boopathi E, Gomes C, Zderic SA, Malkowicz B, Chakrabarti R, Patel DP, Wein AJ, Chacko S (2014) Mechanical stretch upregulates proteins involved in Ca2+ sensitization in urinary bladder smooth muscle hypertrophy. Am J Physiol Cell Physiol 307(6):C542–C553CrossRefPubMedPubMedCentral
11.
go back to reference Tagaya M, Oka M, Ueda M, Takagaki K, Tanaka M, Ohgi T, Yano J (2009) Eviprostat suppresses proinflammatory gene expression in the prostate of rats with partial bladder-outlet obstruction: a genome-wide DNA microarray analysis. Cytokine 47(3):185–193CrossRefPubMed Tagaya M, Oka M, Ueda M, Takagaki K, Tanaka M, Ohgi T, Yano J (2009) Eviprostat suppresses proinflammatory gene expression in the prostate of rats with partial bladder-outlet obstruction: a genome-wide DNA microarray analysis. Cytokine 47(3):185–193CrossRefPubMed
12.
go back to reference Duan LJ, Cao QF, Xu D, Liu HL, Qi J (2017) Bioinformatic analysis of microRNA-mRNA expression profiles of bladder tissue induced by bladder outlet obstruction in a rat model. Mol Med Rep 16(4):4803–4810CrossRefPubMed Duan LJ, Cao QF, Xu D, Liu HL, Qi J (2017) Bioinformatic analysis of microRNA-mRNA expression profiles of bladder tissue induced by bladder outlet obstruction in a rat model. Mol Med Rep 16(4):4803–4810CrossRefPubMed
13.
go back to reference Andersen G, Busso D, Poterszman A, Hwang JR, Wurtz JM, Ripp R, Thierry JC, Egly JM, Moras D (1997) The structure of cyclin H: common mode of kinase activation and specific features. EMBO J 16(5):958–967CrossRefPubMedPubMedCentral Andersen G, Busso D, Poterszman A, Hwang JR, Wurtz JM, Ripp R, Thierry JC, Egly JM, Moras D (1997) The structure of cyclin H: common mode of kinase activation and specific features. EMBO J 16(5):958–967CrossRefPubMedPubMedCentral
14.
go back to reference Jordan P, Cunha C, Carmo-Fonseca M (1997) The cdk7-cyclin H-MAT1 complex associated with TFIIH is localized in coiled bodies. Mol Biol Cell 8(7):1207–1217CrossRefPubMedPubMedCentral Jordan P, Cunha C, Carmo-Fonseca M (1997) The cdk7-cyclin H-MAT1 complex associated with TFIIH is localized in coiled bodies. Mol Biol Cell 8(7):1207–1217CrossRefPubMedPubMedCentral
15.
go back to reference Wang KC, Nguyen P, Weiss A, Yeh YT, Chien HS, Lee A, Teng D, Subramaniam S, Li YS, Chien S (2014) MicroRNA-23b regulates cyclin-dependent kinase-activating kinase complex through cyclin H repression to modulate endothelial transcription and growth under flow. Arterioscler Thromb Vasc Biol 34(7):1437–1445CrossRefPubMedPubMedCentral Wang KC, Nguyen P, Weiss A, Yeh YT, Chien HS, Lee A, Teng D, Subramaniam S, Li YS, Chien S (2014) MicroRNA-23b regulates cyclin-dependent kinase-activating kinase complex through cyclin H repression to modulate endothelial transcription and growth under flow. Arterioscler Thromb Vasc Biol 34(7):1437–1445CrossRefPubMedPubMedCentral
16.
go back to reference Zhang J, Zhu J, Yang L, Guan C, Ni R, Wang Y, Ji L, Tian Y (2015) Interaction with CCNH/CDK7 facilitates CtBP2 promoting esophageal squamous cell carcinoma (ESCC) metastasis via upregulating epithelial-mesenchymal transition (EMT) progression. Tumour Biol 36(9):6701–6714CrossRefPubMed Zhang J, Zhu J, Yang L, Guan C, Ni R, Wang Y, Ji L, Tian Y (2015) Interaction with CCNH/CDK7 facilitates CtBP2 promoting esophageal squamous cell carcinoma (ESCC) metastasis via upregulating epithelial-mesenchymal transition (EMT) progression. Tumour Biol 36(9):6701–6714CrossRefPubMed
17.
go back to reference Singh S, Powell DW, Rane MJ, Millard TH, Trent JO, Pierce WM, Klein JB, Machesky LM, McLeish KR (2003) Identification of the p16-Arc subunit of the Arp 2/3 complex as a substrate of MAPK-activated protein kinase 2 by proteomic analysis. J Biol Chem 278(38):36410–36417CrossRefPubMed Singh S, Powell DW, Rane MJ, Millard TH, Trent JO, Pierce WM, Klein JB, Machesky LM, McLeish KR (2003) Identification of the p16-Arc subunit of the Arp 2/3 complex as a substrate of MAPK-activated protein kinase 2 by proteomic analysis. J Biol Chem 278(38):36410–36417CrossRefPubMed
18.
go back to reference Millard TH, Behrendt B, Launay S, Fütterer K, Machesky LM (2003) Identification and characterisation of a novel human isoform of Arp2/3 complex subunit p16-ARC/ARPC5. Cell Motil Cytoskelet 54(1):81–90CrossRef Millard TH, Behrendt B, Launay S, Fütterer K, Machesky LM (2003) Identification and characterisation of a novel human isoform of Arp2/3 complex subunit p16-ARC/ARPC5. Cell Motil Cytoskelet 54(1):81–90CrossRef
19.
go back to reference Kinoshita T, Nohata N, Watanabe-Takano H, Yoshino H, Hidaka H, Fujimura L, Fuse M, Yamasaki T, Enokida H, Nakagawa M, Hanazawa T, Okamoto Y, Seki N (2012) Actin-related protein 2/3 complex subunit 5 (ARPC5) contributes to cell migration and invasion and is directly regulated by tumor-suppressive microRNA-133a in head and neck squamous cell carcinoma. Int J Oncol 40(6):1770–1778PubMed Kinoshita T, Nohata N, Watanabe-Takano H, Yoshino H, Hidaka H, Fujimura L, Fuse M, Yamasaki T, Enokida H, Nakagawa M, Hanazawa T, Okamoto Y, Seki N (2012) Actin-related protein 2/3 complex subunit 5 (ARPC5) contributes to cell migration and invasion and is directly regulated by tumor-suppressive microRNA-133a in head and neck squamous cell carcinoma. Int J Oncol 40(6):1770–1778PubMed
20.
go back to reference Silverman-Gavrila R, Silverman-Gavrila L, Hou G, Zhang M, Charlton M, Bendeck MP (2011) Rear polarization of the microtubule-organizing center in neointimal smooth muscle cells depends on PKCα, ARPC5, and RHAMM. Am J Pathol 178(2):895–910CrossRefPubMedPubMedCentral Silverman-Gavrila R, Silverman-Gavrila L, Hou G, Zhang M, Charlton M, Bendeck MP (2011) Rear polarization of the microtubule-organizing center in neointimal smooth muscle cells depends on PKCα, ARPC5, and RHAMM. Am J Pathol 178(2):895–910CrossRefPubMedPubMedCentral
21.
go back to reference Gournier H, Goley ED, Niederstrasser H, Trinh T, Welch MD (2001) Reconstitution of human Arp2/3 complex reveals critical roles of individual subunits in complex structure and activity. Mol Cell 8(5):1041–1052CrossRefPubMed Gournier H, Goley ED, Niederstrasser H, Trinh T, Welch MD (2001) Reconstitution of human Arp2/3 complex reveals critical roles of individual subunits in complex structure and activity. Mol Cell 8(5):1041–1052CrossRefPubMed
23.
go back to reference Engels BM, Hutvagner G (2006) Principles and effects of microRNA mediated post-transcriptional gene regulation. Oncogene 25(46):6163–6169CrossRefPubMed Engels BM, Hutvagner G (2006) Principles and effects of microRNA mediated post-transcriptional gene regulation. Oncogene 25(46):6163–6169CrossRefPubMed
26.
go back to reference Calin GA, Croce CM (2006) MicroRNAs and chromosomal abnormalities in cancer cells. Oncogene 25(46):6202–6210CrossRefPubMed Calin GA, Croce CM (2006) MicroRNAs and chromosomal abnormalities in cancer cells. Oncogene 25(46):6202–6210CrossRefPubMed
27.
go back to reference Muys BR, Lorenzi JC, Zanette DL, Lima e Bueno Rde B, de Araújo LF, Dinarte-Santos AR, Alves CP, Ramão A, de Molfetta GA, Vidal DO, Silva WA Jr (2016) Placenta-Enriched LincRNAs MIR503HG and LINC00629 Decrease migration and invasion potential of JEG-3 cell line. PLoS One 11(3):e0151560CrossRefPubMedPubMedCentral Muys BR, Lorenzi JC, Zanette DL, Lima e Bueno Rde B, de Araújo LF, Dinarte-Santos AR, Alves CP, Ramão A, de Molfetta GA, Vidal DO, Silva WA Jr (2016) Placenta-Enriched LincRNAs MIR503HG and LINC00629 Decrease migration and invasion potential of JEG-3 cell line. PLoS One 11(3):e0151560CrossRefPubMedPubMedCentral
28.
go back to reference Brandenberger R, Wei H, Zhang S, Lei S, Murage J, Fisk GJ, Li Y, Xu C, Fang R, Guegler K, Rao MS, Mandalam R, Lebkowski J, Stanton LW (2004) Transcriptome characterization elucidates signaling networks that control human ES cell growth and differentiation. Nat Biotechnol 22(6):707–716CrossRefPubMed Brandenberger R, Wei H, Zhang S, Lei S, Murage J, Fisk GJ, Li Y, Xu C, Fang R, Guegler K, Rao MS, Mandalam R, Lebkowski J, Stanton LW (2004) Transcriptome characterization elucidates signaling networks that control human ES cell growth and differentiation. Nat Biotechnol 22(6):707–716CrossRefPubMed
Metadata
Title
Identification of key genes and construction of microRNA–mRNA regulatory networks in bladder smooth muscle cell response to mechanical stimuli using microarray expression profiles and bioinformatics analysis
Authors
Liao Peng
De-Yi Luo
Publication date
01-02-2018
Publisher
Springer Berlin Heidelberg
Published in
World Journal of Urology / Issue 2/2018
Print ISSN: 0724-4983
Electronic ISSN: 1433-8726
DOI
https://doi.org/10.1007/s00345-017-2132-3

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