miR-141 regulates TGF-β1-induced epithelial-mesenchymal transition through repression of HIPK2 expression in renal tubular epithelial cells

Huang,Yuanhang; Tong,Junrong; He,Feng; Yu,Xinpei; Fan,Liming; Hu,Jing; Tan,Jiangping; Chen,Zhengliang

doi:10.3892/ijmm.2014.2008

miR-141 regulates TGF-β1-induced epithelial-mesenchymal transition through repression of HIPK2 expression in renal tubular epithelial cells

Authors:
- Yuanhang Huang
- Junrong Tong
- Feng He
- Xinpei Yu
- Liming Fan
- Jing Hu
- Jiangping Tan
- Zhengliang Chen
View Affiliations

Affiliations: Department of Immunology, School of Basic Medical Sciences, Southern Medical University, Guangzhou, Guangdong 510515, P.R. China, Department of Nephrology, Guanzhou General Hospital of Guanzhou Military Command, Guangzhou, Guangdong 510010, P.R. China, Geriatric Infection and Organ Function Support Laboratory, Guanzhou General Hospital of Guanzhou Military Command, Guangzhou, Guangdong 510010, P.R. China
Published online on: November 24, 2014 https://doi.org/10.3892/ijmm.2014.2008
Pages: 311-318
Copyright: © Huang et al. This is an open access article distributed under the terms of Creative Commons Attribution License [CC BY_NC 3.0].

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Epithelial‑mesenchymal transition (EMT) plays a critical role in embryonic development, wound healing, tissue regeneration, cancer progression and organ fibrosis. The proximal tubular epithelial cells undergo EMT, resulting in matrix‑producing fibroblasts and thereby contribute to the pathogenesis of renal fibrosis. The profibrotic cytokine, TGF‑β, is now recognized as the main pathogenic driver that has been shown to induce EMT in tubular epithelial cells. Increasing evidence indicate that HIPK2 dysfunction may play a role in fibroblasts behavior, and therefore, HIPK2 may be considered as a novel potential target for anti‑fibrosis therapy. Recently, members of the miR‑200 family (miR‑200a, b and c and miR‑141) have been shown to inhibit EMT. However, the steps of the multifactorial renal fibrosis progression that these miRNAs regulate, particularly miR‑141, are unclear. To study the functional importance of miR‑141 in EMT, a well‑established in vitro EMT assay was used to demonstrate renal tubulointerstitial fibrosis; transforming growth factor‑β1‑induced EMT in HK‑2 cells. Overexpression of miR‑141 in HK‑2 cells, either with or without TGF‑β1 treatment, hindered EMT by enhancing E‑cadherin and decreasing vimentin and fibroblast‑specific protein 1 expression. miR‑141 expression was repressed during EMT in a dose‑ and time‑dependent manner through upregulation of HIPK2 expression. Ectopic expression of HIPK2 promoted EMT by decreasing E‑cadherin. Furthermore, co‑transfection of miR‑141 with the HIPK2 ORF clone partially inhibited EMT by restoring E‑cadherin expression. miR‑141 downregulated the expression of HIPK2 via direct interaction with the 3'‑untranslated region of HIPK2. Taken together, these findings aid in the understanding of the role and mechanism of miR‑141 in regulating renal fibrosis via the TGF‑β1/miR‑141/HIPK2/EMT axis, and miR‑141 may represent novel biomarkers and therapeutic targets in the treatment of renal fibrosis.

View Figures

View References

1	Liu Y: Epithelial to mesenchymal transition in renal fibrogenesis: pathologic significance, molecular mechanism, and therapeutic intervention. J Am Soc Nephrol. 15:1–12. 2004. View Article : Google Scholar
2	Zeisberg M and Kalluri R: The role of epithelial-to-mesenchymal transition in renal fibrosis. J Mol Med (Berl). 82:175–181. 2004. View Article : Google Scholar
3	Kalluri R and Weinberg RA: The basics of epithelial-mesenchymal transition. J Clin Invest. 119:1420–1428. 2009. View Article : Google Scholar : PubMed/NCBI
4	Strutz F, Okada H, Lo CW, Danoff T, Carone RL, Tomaszewski JE and Neilson EG: Identification and characterization of a fibroblast marker: FSP1. J Cell Biol. 130:393–405. 1995. View Article : Google Scholar : PubMed/NCBI
5	Strutz F and Müller GA: Renal fibrosis and the origin of the renal fibroblast. Nephrol Dial Transplant. 21:3368–3370. 2006. View Article : Google Scholar : PubMed/NCBI
6	Liu Y: New insights into epithelial-mesenchymal transition in kidney fibrosis. J Am Soc Nephrol. 21:212–222. 2010. View Article : Google Scholar
7	Li Y, Yang J, Luo JH, Dedhar S and Liu Y: Tubular epithelial cell dedifferentiation is driven by the helix-loop-helix transcriptional inhibitor id1. J Am Soc Nephrol. 18:449–460. 2007. View Article : Google Scholar : PubMed/NCBI
8	Tan R, Zhang J, Tan X, Zhang X, Yang J and Liu Y: Downregulation of SnoN expression in obstructive nephropathy is mediated by an enhanced ubiquitin-dependent degradation. J Am Soc Nephrol. 17:2781–2791. 2006. View Article : Google Scholar : PubMed/NCBI
9	Yang J, Shultz RW, Mars WM, Wegner RE, Li Y, Dai C, Nejak K and Liu Y: Disruption of tissue-type plasminogen activator gene in mice reduces renal interstitial fibrosis in obstructive nephropathy. J Clin Invest. 110:1525–1538. 2002. View Article : Google Scholar : PubMed/NCBI
10	Yang J, Zhang X, Li Y and Liu Y: Downregulation of smad transcriptional corepressors snon and ski in the fibrotic kidney: an amplification mechanism for TGF-beta1 signaling. J Am Soc Nephrol. 14:3167–3177. 2003. View Article : Google Scholar : PubMed/NCBI
11	Lan HY: Diverse roles of TGF-beta/smads in renal fibrosis and inflammation. Int J Biol Sci. 7:1056–1067. 2011. View Article : Google Scholar
12	Meng XM, Chung AC and Lan HY: Role of the TGF-beta/BMP-7/smad pathways in renal diseases. Clin Sci (Lond). 124:243–254. 2013. View Article : Google Scholar
13	Kalluri R and Neilson EG: Epithelial-mesenchymal transition and its implications for fibrosis. J Clin Invest. 112:1776–1784. 2003. View Article : Google Scholar : PubMed/NCBI
14	Branton MH and Kopp JB: TGF-beta and fibrosis. Microbes Infect. 1:1349–1365. 1999. View Article : Google Scholar : PubMed/NCBI
15	Patel V and Noureddine L: MicroRNAs and fibrosis. Curr Opin Nephrol Hypertens. 21:410–416. 2012. View Article : Google Scholar : PubMed/NCBI
16	Bartel DP: MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI
17	Wienholds E, Kloosterman WP, Miska E, Alvarez-Saavedra E, Berezikov E, de Bruijn E, Horvitz HR, Kauppinen S and Plasterk RH: MicroRNA expression in zebrafish embryonic development. Science. 309:310–311. 2005. View Article : Google Scholar : PubMed/NCBI
18	Yi R, O’Carroll D, Pasolli HA, Zhang Z, Dietrich FS, Tarakhovsky A and Fuchs E: Morphogenesis in skin is governed by discrete sets of differentially expressed microRNAs. Nat Genet. 38:356–362. 2006. View Article : Google Scholar : PubMed/NCBI
19	Esquela-Kerscher A and Slack FJ: Oncomirs - microRNAs with a role in cancer. Nat Rev. Cancer. 6:259–269. 2006.
20	Zarjou A, Yang S, Abraham E, Agarwal A and Liu G: Identification of a microRNA signature in renal fibrosis: role of miR-21. Am J Physiol Renal Physiol. 301:F793–F801. 2011. View Article : Google Scholar : PubMed/NCBI
21	Xiong M, Jiang L, Zhou Y, Qiu W, Fang L, Tan R, Wen P and Yang J: The miR-200 family regulates TGF-beta1-induced renal tubular epithelial to mesenchymal transition through smad pathway by targeting ZEB1 and ZEB2 expression. Am J Physiol Renal Physiol. 302:F369–F379. 2012. View Article : Google Scholar
22	Wang B, Komers R, Carew R, Winbanks CE, Xu B, Herman-Edelstein M, Koh P, Thomas M, Jandeleit-Dahm K, Gregorevic P, Cooper ME and Kantharidis P: Suppression of microRNA-29 expression by TGF-beta1 promotes collagen expression and renal fibrosis. J Am Soc Nephrol. 23:252–265. 2012. View Article : Google Scholar :
23	Korpal M, Lee ES, Hu G and Kang Y: The miR-200 family inhibits epithelial-mesenchymal transition and cancer cell migration by direct targeting of E-cadherin transcriptional repressors ZEB1 and ZEB2. J Biol Chem. 283:14910–14914. 2008. View Article : Google Scholar : PubMed/NCBI
24	Calzado MA, Renner F, Roscic A and Schmitz ML: HIPK2: a versatile switchboard regulating the transcription machinery and cell death. Cell Cycle. 6:139–143. 2007. View Article : Google Scholar : PubMed/NCBI
25	Lee W, rews BC, Faust M, Walldorf U and Verheyen EM: Hipk is an essential protein that promotes notch signal transduction in the drosophila eye by inhibition of the global co-repressor groucho. Dev Biol. 325:263–272. 2009. View Article : Google Scholar
26	Lee W, Swarup S, Chen J, Ishitani T and Verheyen EM: Homeodomain-interacting protein kinases (Hipks) promote Wnt/Wg signaling through stabilization of beta-catenin/Arm and stimulation of target gene expression. Development. 136:241–251. 2009. View Article : Google Scholar
27	Zhang J, Pho V, Bonasera SJ, Holtzman J, Tang AT, Hellmuth J, Tang S, Janak PH, Tecott LH and Huang EJ: Essential function of HIPK2 in TGFbeta-dependent survival of midbrain dopamine neurons. Nat Neurosci. 10:77–86. 2007. View Article : Google Scholar
28	D’Orazi G, Cecchinelli B, Bruno T, Manni I, Higashimoto Y, Saito S, Gostissa M, Coen S, Marchetti A, Del Sal G, Piaggio G, Fanciulli M, Appella E and Soddu S: Homeodomain-interacting protein kinase-2 phosphorylates p53 at ser 46 and mediates apoptosis. Nat Cell Biol. 4:11–19. 2002. View Article : Google Scholar : PubMed/NCBI
29	Hofmann TG, Möller A, Sirma H, Zentgraf H, Taya Y, Dröge W, Will H and Schmitz ML: Regulation of p53 activity by its interaction with homeodomain-interacting protein kinase-2. Nat Cell Biol. 4:1–10. 2002. View Article : Google Scholar
30	Rinaldo C, Prodosmo A, Siepi F and Soddu S: HIPK2: a multitalented partner for transcription factors in DNA damage response and development. Biochem Cell Biol. 85:411–418. 2007. View Article : Google Scholar : PubMed/NCBI
31	Jin Y, Ratnam K, Chuang PY, Fan Y, Zhong Y, Dai Y, Mazloom AR, Chen EY, D’Agati V, Xiong H, Ross MJ, Chen N, Ma’ayan A and He JC: A systems approach identifies HIPK2 as a key regulator of kidney fibrosis. Nat Med. 18:580–588. 2012. View Article : Google Scholar : PubMed/NCBI
32	Ricci A, Cherubini E, Ulivieri A, Lavra L, Sciacchitano S, Scozzi D, Mancini R, Ciliberto G, Bartolazzi A, Bruno P, Graziano P and Mariotta S: Homeodomain-interacting protein kinase2 in human idiopathic pulmonary fibrosis. J Cell Physiol. 228:235–241. 2013. View Article : Google Scholar
33	Wang B, Koh P, Winbanks C, Coughlan MT, McClelland A, Watson A, Jandeleit-Dahm K, Burns WC, Thomas MC, Cooper ME and Kantharidis P: miR-200a prevents renal fibrogenesis through repression of TGF-beta2 expression. Diabetes. 60:280–287. 2011. View Article : Google Scholar :
34	Tamagawa S, Beder LB, Hotomi M, Gunduz M, Yata K, Grenman R and Yamanaka N: Role of miR-200c/mir-141 in the regulation of epithelial-mesenchymal transition and migration in head and neck squamous cell carcinoma. Int J Mol Med. 33:879–886. 2014.PubMed/NCBI
35	Wallace K, Burt AD and Wright MC: Liver fibrosis. Biochem J. 411:1–18. 2008. View Article : Google Scholar : PubMed/NCBI
36	Wynn TA: Integrating mechanisms of pulmonary fibrosis. J Exp Med. 208:1339–1350. 2011. View Article : Google Scholar : PubMed/NCBI
37	Yang J and Liu Y: Dissection of key events in tubular epithelial to myofibroblast transition and its implications in renal interstitial fibrosis. Am J Pathol. 159:1465–1475. 2001. View Article : Google Scholar : PubMed/NCBI
38	Gregory PA, Bert AG, Paterson EL, Barry SC, Tsykin A, Farshid G, Vadas MA, Khew-Goodall Y and Goodall GJ: The mir-200 family and miR-205 regulate epithelial to mesenchymal transition by targeting ZEB1 and SIP1. Nat Cell Biol. 10:593–601. 2008. View Article : Google Scholar : PubMed/NCBI
39	Hill C, Flyvbjerg A, Gronbaek H, Petrik J, Hill DJ, Thomas CR, Sheppard MC and Logan A: The renal expression of transforming growth factor-beta isoforms and their receptors in acute and chronic experimental diabetes in rats. Endocrinology. 141:1196–1208. 2000.PubMed/NCBI
40	Chalazonitis A, Tang AA, Shang Y, Pham TD, Hsieh I, Setlik W, Gershon MD and Huang EJ: Homeodomain interacting protein kinase 2 regulates postnatal development of enteric dopaminergic neurons and glia via BMP signaling. J Neurosci. 31:13746–13757. 2011. View Article : Google Scholar : PubMed/NCBI
41	Harada J, Kokura K, Kanei-Ishii C, Nomura T, Khan MM, Kim Y and Ishii S: Requirement of the co-repressor homeodomain-interacting protein kinase 2 for ski-mediated inhibition of bone morphogenetic protein-induced transcriptional activation. J Biol Chem. 278:38998–39005. 2003. View Article : Google Scholar : PubMed/NCBI
42	Hofmann TG, Stollberg N, Schmit ML and Will H: HIPK2 regulates transforming growth factor-beta-induced c-jun NH (2)-terminal kinase activation and apoptosis in human hepatoma cells. Cancer Res. 63:8271–8277. 2003.PubMed/NCBI