Radiation and Environmental Biophysics

, Volume 57, Issue 1, pp 41–54 | Cite as

Regulation of type I collagen expression by microRNA-29 following ionizing radiation

  • Hiroyuki YanoEmail author
  • Ryoji Hamanaka
  • Miki Nakamura-Ota
  • Juan Juan Zhang
  • Noritaka Matsuo
  • Hidekatsu Yoshioka
Original Article


Radiation-induced fibrosis (RIF) is thought to involve the excessive accumulation of collagen and other extracellular matrix components; previously, we reported that ionizing radiation increased the type I collagen expression and that transforming growth factor (TGF)-β was involved in this increase through activating its downstream mediator, Smad3. A recent study found that microRNAs (miRNAs)—small, noncoding sequences approximately 20 nucleotides long—negatively regulate the gene expression posttranscriptionally, and it has been suggested that miRNAs play essential roles in cellular processes, including fibrosis. However, their role in the development of RIF remains unexplored. In the present study, we examined the effects of miRNA on the expression of type I collagen induced by ionizing radiation and the mechanisms underlying the miRNA expression observed following ionizing radiation. We analyzed the regulation of miRNA following ionizing radiation by an miRNA real-time PCR, and found that miR-29 family members were downregulated in irradiated mouse fibroblasts and directly targeted type I collagen genes by specifically binding to the 3ʹ untranslated region. We also found that the overexpression of miR-29 inhibited the ionizing radiation-induced expression of type I collagen, whereas the knockdown of miR-29 enhanced it. In addition, TGF-β/Smad-signaling significantly decreased the transcription of miR-29, whereas the inhibition of this signaling pathway cancelled this decrease. In conclusion, miR-29 was involved in the regulation of type I collagen expression through the TGF-β/Smad-signaling pathway in irradiated cells, suggesting that miR-29 may be an important regulator of RIF.


Ionizing radiation Type I collagen MicroRNA Fibrosis 



Radiation-induced fibrosis



CHIP assay

Chromatin immunoprecipitation assay


Extracellular matrix



We thank Mr. K. Kai, Ms. H. Sato, and the staff members of the Department of Matrix Medicine, Faculty of Medicine, Oita University. This work was supported by Grant-in-Aid for Young Scientists (No. 15K19697 to H. Y.) from the Ministry of Education, Culture, Sports, Science, and Technology of Japan.

Supplementary material

411_2017_723_MOESM1_ESM.docx (21 kb)
Supplementary material 1 (DOCX 21 KB)


  1. Agarwal V, Bell GW, Nam JW, Bartel DP (2015) Predicting effective microRNA target sites in mammalian mRNAs. Elife 12:1–38Google Scholar
  2. Bartel DP (2014) MicroRNAs: genomics, biogenesis, mechanism, and function. Cell 116:281–297CrossRefGoogle Scholar
  3. Border WA, Noble NA (1994) Transforming growth factor b in tissue fibrosis. N Engl J Med 331:1286–1292CrossRefGoogle Scholar
  4. Brennecke J, Hipfner DR, Stark A, Russell RB, Cohen SM (2003) bantam encodes a developmentally regulated microRNA that controls cell proliferation and regulates the proapoptotic gene hid in Drosophila. Cell 113:25–36CrossRefGoogle Scholar
  5. Brown BD, Naldini L (2009) Exploiting and antagonizing microRNA regulation for therapeutic and experimental applications. Nat Rev Genet 10:578–585CrossRefGoogle Scholar
  6. Chang TC, Wentzel EA, Kent OA, Ramachandran K, Mullendore M, Lee KH et al (2007) Transactivation of miR-34a by p53 broadly influences gene expression and promotes apoptosis. Mol Cell 26:745–752CrossRefGoogle Scholar
  7. Chang TC, Yu D, Lee YS, Wentzel EA, Arking DE, West KM et al (2008) Widespread microRNA repression by Myc contributes to tumorigenesis. Nat Genet 40:43–50CrossRefGoogle Scholar
  8. Chen CZ, Li L, Lodish HF, Bartel DP (2004) MicroRNAs modulate hematopoietic lineage differentiation. Science 303:83–86ADSCrossRefGoogle Scholar
  9. Cushing L, Kuang PP, Qian J, Shao F, Wu J, Little F et al (2011) MIR-29 is a major regulator of genes associated with pulmonary fibrosis. Am J Respir Cell Mol Biol 45:287–294CrossRefGoogle Scholar
  10. Davis BN, Hilyard AC, Lagna G, Hata A (2008) SMAD proteins control DROSHA-mediated microRNA maturation. Nature 454:56–61ADSCrossRefGoogle Scholar
  11. Elnakish MT, Kuppusamy P, Khan M (2013) Stem cell transplantation as a therapy for cardiac fibrosis. J Pathol 229:347–354CrossRefGoogle Scholar
  12. Friedman SL (2007) Liver fibrosis: from mechanisms to treatment. Gastroenterol Clin Biol 31:812–814CrossRefGoogle Scholar
  13. Gordon MK, Hahn RA (2010) Collagens. Cell Tissue Res 339:247–257CrossRefGoogle Scholar
  14. Iredale JP, Thompson A, Henderson NC (2013) Extracellular matrix degradation in liver fibrosis. Biochem Biophys Acta 1832:876–883Google Scholar
  15. Kurinna S, Schäfer M, Ostano P, Karouzakis E, Chiorino G, Bloch W et al (2014) A novel Nrf2-miR-29-desmocollin-2 axis regulates desmosome function in keratinocytes. Nat Commun 5:5099CrossRefGoogle Scholar
  16. Lee EL, Baek M, Gusev Y, Brackett DJ, Nuovo GJ, Schmittgen TD (2008) Systematic evaluation of microRNA processing patterns in tissues, cell lines, and tumors. RNA 14:35–42CrossRefGoogle Scholar
  17. Liu G, Friggeri A, Yang Y, Milosevic J, Ding Q, Thannickal VJ et al (2010) miR-21 mediates fibrogenic activation of pulmonary fibroblasts and lung fibrosis. J Exp Med 207:1589–1597CrossRefGoogle Scholar
  18. Lorenzen JM, Haller H, Thum T (2011) MicroRNAs as mediators and therapeutic targets in chronic kidney disease. Nat Rev Nephrol 286:286–294CrossRefGoogle Scholar
  19. Maher TM, Wells AU, Laurent GJ (2007) Idiopathic pulmonary fibrosis: multiple causes and multiple mechanisms? Eur Respir J 30:835–839CrossRefGoogle Scholar
  20. Martin M, Lefaix J, Delanian S (2000) TGF-β and radiation fibrosis: a master switch and a specific therapeutic target ? Int J Radiat Oncol Biol Phys 47:277–290CrossRefGoogle Scholar
  21. Maurer B, Stanczyk J, Jungel A, Akhmetshina A, Trenkmann M, Brock M et al (2010) MicroRNA-29, a key regulator of collagen expression in systemic sclerosis. Arthritis Rheum 62:1733–1743CrossRefGoogle Scholar
  22. Noetel A, Kwiecinski M, Elfimova N, Huang J, Odenthal M (2012) microRNA are central players in anti- and profibrotic gene regulation during liver fibrosis. Front Physiol 19:1–6Google Scholar
  23. O’Donnell KA, Wentzel EA, Zeller KI, Dang CV, Mendell JT (2005) c-Myc regulated microRNAs modulate E2F1 expression. Nature 435:839–843ADSCrossRefGoogle Scholar
  24. Pandit KV, Corcoran D, Yousef H, Yarlagadda M, Tzouvelekis A, Gibson KF et al (2010) Inhibition and role of let-7d in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med 182:220–229CrossRefGoogle Scholar
  25. Pohlers D, Brenmoehl J, Löffler I, Müller CK, Leipner C, Schultze-Mosgau S et al (2009) TGF-β and fibrosis in different organs—molecular pathway imprints. Biochim Biophys Acta 1792:746–756CrossRefGoogle Scholar
  26. Qin W, Chung AC, Huang XR, Meng XM, Hui DS, Yu CM et al (2011) TGF-β/Smad3 signaling promotes renal fibrosis by inhibiting miR-29. J Am Soc Nephrol 22:1462–1474CrossRefGoogle Scholar
  27. Raver-Shapira N, Marciano E, Meiri E, Spector Y, Rosenfeld N, Moskovits N et al (2007) Transcriptional activation of miR-34a contributes to p53-mediated apoptosis. Mol Cell 26:731–743CrossRefGoogle Scholar
  28. Rodemann HP, Bamberg M (1995) Cellular basis of radiation-induced fibrosis. Radiother Oncol 32:83–90CrossRefGoogle Scholar
  29. Roderburg C, Urban GW, Bettermann K, Vucur M, Zimmermann H, Schmidt S et al (2011) Micro-RNA profiling reveals a role for miR-29 in human and murine liver fibrosis. Hepatology 53:209–218CrossRefGoogle Scholar
  30. Rødningen OK, Børresen-Dale AL, Alsner J, Hastie T, Overgaard J (2008) Radiation induced gene expression in human subcutaneous fibroblasts is predictive of radiation-induced fibrosis. Radiother Oncol 86:314–320CrossRefGoogle Scholar
  31. Schratt GM, Tuebing F, Nigh EA, Kane CG, Sabatini ME, Kiebler M et al (2006) A brain-specific microRNA regulates dendritic spine development. Nature 439:283–289ADSCrossRefGoogle Scholar
  32. Slack JL, Liska DJ, Bornstein P (1993) Regulation of expression of the type I collagen genes. Am J Med Genet 45:140–151CrossRefGoogle Scholar
  33. Sugihara T, Murano M, Tanaka K, Oghiso Y (2008) Inverse dose-rate-effects on the expressions of extra-cellular matrix-related genes in low-dose-rate g-ray irradiated murine cells. J Radiat Res 49:231–240CrossRefGoogle Scholar
  34. Tarasov V, Jung P, Verdoodt B, Lodygin D, Epanchintsev A, Menssen A et al (2007) Differential regulation of microRNAs by p53 revealed by massively parallel sequencing: miR-34a is a p53 target that induces apoptosis and G1-arrest. Cell Cycle 6:1586–1593CrossRefGoogle Scholar
  35. van Rooij E, Sutherland LB, Thatcher JE, DiMaio JM, Naseem RH, Marshall WS et al (2008) Dysregulation of microRNAs after myocardial infarction reveals a role of miR-29 in cardiac fibrosis. Proc Natl Acad Sci 105:13027–13032ADSCrossRefGoogle Scholar
  36. van der Rest M, Garrone R (1991) Collagen family of proteins. FASEB J 5:2814–2823CrossRefGoogle Scholar
  37. Wang H, Garzon R, Sun H, Ladner KJ, Singh R, Dahlman J et al (2008) NF-kappaB-YY1-miR-29 regulatory circuitry in skeletal myogenesis and rhabdomyosarcoma. Cancer Cell 14:369–381CrossRefGoogle Scholar
  38. Yang L, Cheng P, Chen C, He HB, Xie GQ, Zhou HD et al (2012) miR-93/Sp7 function loop mediates osteoblast mineralization. J Bone Miner Res 27:1598–1606CrossRefGoogle Scholar
  39. Yano H, Hamanaka R, Nakamura M, Sumiyoshi H, Matsuo N, Yoshioka H (2012) Smad, but not MAPK, pathway mediates the expression of type I collagen in radiation induced fibrosis. BBRC 418:457–463Google Scholar
  40. Zhou L, Wang L, Lu L, Jiang P, Sun H, Wang H (2012) Inhibition of miR-29 by TGF-beta-Smad3 signaling through dual mechanisms promotes transdifferentiation of mouse myoblasts into myofibroblasts. Plos One 7:e33766ADSCrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2017

Authors and Affiliations

  • Hiroyuki Yano
    • 1
    Email author
  • Ryoji Hamanaka
    • 2
    • 3
  • Miki Nakamura-Ota
    • 2
  • Juan Juan Zhang
    • 4
  • Noritaka Matsuo
    • 4
  • Hidekatsu Yoshioka
    • 4
    • 5
  1. 1.Research Promotion InstituteOita UniversityYufuJapan
  2. 2.Department of Cell Biology, Faculty of MedicineOita UniversityYufuJapan
  3. 3.Department of Human SciencesOita University of Nursing and Human SciencesOitaJapan
  4. 4.Department of Matrix Medicine, Faculty of MedicineOita UniversityYufuJapan
  5. 5.Department of Clinical ExaminationShinbeppu HospitalBeppuJapan

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