Archives of Dermatological Research

, Volume 305, Issue 1, pp 9–15 | Cite as

microRNA-7 down-regulation mediates excessive collagen expression in localized scleroderma

  • Mitsuhiko Etoh
  • Masatoshi Jinnin
  • Katsunari Makino
  • Keitaro Yamane
  • Wakana Nakayama
  • Jun Aoi
  • Noritoshi Honda
  • Ikko Kajihara
  • Takamitsu Makino
  • Satoshi Fukushima
  • Hironobu Ihn
Original Paper

Abstract

Localized scleroderma (LSc), a connective tissue disorder restricted to the skin and subcutaneous tissue, is characterized by skin fibrosis due to an excessive deposition of types I collagen. The mechanism of such fibrosis is still unknown, but epigenetics may play some roles in the excessive collagen expression. In the present study, we investigated the mechanism of fibrosis seen in LSc, focusing on microRNA (miRNA). miRNA expression was determined by PCR array, real-time PCR, and in situ hybridization. The function of miRNA was evaluated using specific inhibitor. Immunoblotting was performed to detect α2(I) collagen protein. PCR array analysis using tissue miRNA demonstrated miR-7 level was significantly decreased in LSc skin as well as keloid tissue compared to normal skin in vivo. In situ hybridization also showed miR-7 expression in dermal fibroblasts was decreased in LSc dermis. The transfection of specific inhibitor for miR-7 into cultured normal dermal fibroblasts resulted in the up-regulation of α2(I) collagen protein in vitro. Also, the serum levels of miR-7 were significantly decreased in LSc patients compared with healthy controls, but serum miR-29a levels not. Systemic or local down-regulation of miR-7 may contribute to the pathogenesis of LSc via the overexpression of α2(I) collagen, and serum miR-7 may be useful as a disease marker. Investigation of the regulatory mechanisms of LSc by miRNA may lead to new treatments by the transfection into the lesional skin of this disease.

Keywords

Collagen Epigenetics Fibrosis 

Notes

Acknowledgments

We thank Ms. Junko Suzuki and Ms. Chiemi Shiotsu for their valuable technical assistance. This study was supported in part by a grant for scientific research from the Japanese Ministry of Education, Science, Sports and Culture, by project research on intractable diseases from the Japanese Ministry of Health, Labour and Welfare, and by Shiseido Research Grant.

References

  1. 1.
    Arkachaisri T, Vilaiyuk S, Li S et al (2009) The localized scleroderma skin severity index and physician global assessment of disease activity: a work in progress toward development of localized scleroderma outcome measures. J Rheumatol 36:2819–2829PubMedCrossRefGoogle Scholar
  2. 2.
    Arkachaisri T, Vilaiyuk S, Torok KS, Medsger TA (2010) Development and initial validation of the localized scleroderma skin damage index and physician global assessment of disease damage: a proof-of-concept study. Rheumatology (Oxford) 49:373–381CrossRefGoogle Scholar
  3. 3.
    Asano Y, Ihn H, Jinnin M, Mimura Y, Tamaki K (2006) Involvement of αvβ5 integrin in the establishment of autocrine TGF-β signaling in dermal fibroblasts derived from localized scleroderma. J Invest Dermatol 126:1761–1769PubMedCrossRefGoogle Scholar
  4. 4.
    Chen H, Shalom-Feuerstein R, Riley J et al (2010) miR-7 and miR-214 are specifically expressed during neuroblastoma differentiation, cortical development and embryonic stem cells differentiation, and control neurite outgrowth in vitro. Biochem Biophys Res Commun 394:921–927PubMedCrossRefGoogle Scholar
  5. 5.
    Cohly HP, Scott H, Ndebele K, Jenkins JK, Angel MF (2002) Differential gene expression of fibroblasts: keloid versus normal. Int J Mol Sci 3:1162–1176CrossRefGoogle Scholar
  6. 6.
    Falanga V, Medsger TJ, Reichlin M (1985) Anti-nuclear and anti-single-stranded DNA in linear scleroderma. Arch Dermatol 123:350–353CrossRefGoogle Scholar
  7. 7.
    Furer V, Greenberg JD, Attur M, Abramson SB, Pillinger MH (2010) The role of microRNA in rheumatoid arthritis and other autoimmune diseases. Clin Immunol 136:1–15PubMedCrossRefGoogle Scholar
  8. 8.
    Ihn H, Sato S, Fujimoto M, Kikuchi K, Takehara K (1996) Clinical significance of serum levels of soluble interleukin-2 receptor in patients with localized scleroderma. Br J Dermatol 134:843–847PubMedCrossRefGoogle Scholar
  9. 9.
    Jiang L, Liu X, Chen Z et al (2010) MicroRNA-7 targets IGF1R (insulin-like growth factor 1 receptor) in tongue squamous cell carcinoma cells. Biochem J 432:199–205PubMedCrossRefGoogle Scholar
  10. 10.
    Kanemaru H, Fukushima S, Yamashita J et al (2011) The circulating microRNA-221 level in patients with malignant melanoma as a new tumor marker. J Dermatol Sci 61:187–193PubMedCrossRefGoogle Scholar
  11. 11.
    Kubo M, Ihn H, Yamane K, Tamaki K (2001) Up-regulated expression of transforming growth factor β receptors in dermal fibroblasts in skin sections from patients with localized scleroderma. Arthritis Rheum 44:731–734PubMedCrossRefGoogle Scholar
  12. 12.
    Li J, Zhang Y, Kuruba R et al (2011) Roles of microRNA-29a in the antifibrotic effect of farnesoid X receptor in hepatic stellate cells. Mol Pharmacol 80:191–200PubMedCrossRefGoogle Scholar
  13. 13.
    Long JM, Lahiri DK (2011) MicroRNA-101 downregulates Alzheimer’s amyloid-β precursor protein levels in human cell cultures and is differentially expressed. Biochem Biophys Res Commun 404:889–895PubMedCrossRefGoogle Scholar
  14. 14.
    Makino T, Jinnin M, Muchemwa FC et al (2010) Basic fibroblast growth factor stimulates the proliferation of human dermal fibroblasts via the ERK1/2 and JNK pathways. Br J Dermatol 162:717–723PubMedCrossRefGoogle Scholar
  15. 15.
    Maurer B, Stanczyk J, Jüngel A et al (2010) MicroRNA-29, a key regulator of collagen expression in systemic sclerosis. Arthritis Rheum 62:1733–1743PubMedCrossRefGoogle Scholar
  16. 16.
    Pandit KV, Milosevic J, Kaminski N (2011) MicroRNAs in idiopathic pulmonary fibrosis. Transl Res 157:191–199PubMedCrossRefGoogle Scholar
  17. 17.
    Sato S, Ihn H, Soma Y et al (1993) Antihistone antibodies in patients with localized scleroderma. Arthritis Rheum 36:1137–1141PubMedCrossRefGoogle Scholar
  18. 18.
    Sato S, Fujimoto M, Ihn H, Kikuchi K, Takehara K (1994) Clinical characteristics associated with antihistone antibodies in patients with localized scleroderma. J Am Acad Dermatol 31:567–571PubMedCrossRefGoogle Scholar
  19. 19.
    Takehara K, Kikuchi K, Soma Y, Igarashi A, Ishibashi Y (1990) Anti-single-stranded DNA antibody and muscle involvement in localized scleroderma. Arch Dermatol 126:1368PubMedCrossRefGoogle Scholar
  20. 20.
    Uziel Y, Krafchik B, Feldman B, Silverman E, Rubin L, Laxer R (1994) Serum levels of soluble interleukin-2 receptor. A marker of disease activity in localized scleroderma. Arthritis Rheum 37:898–901PubMedCrossRefGoogle Scholar

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Mitsuhiko Etoh
    • 1
  • Masatoshi Jinnin
    • 1
  • Katsunari Makino
    • 1
  • Keitaro Yamane
    • 1
  • Wakana Nakayama
    • 1
  • Jun Aoi
    • 1
  • Noritoshi Honda
    • 1
  • Ikko Kajihara
    • 1
  • Takamitsu Makino
    • 1
  • Satoshi Fukushima
    • 1
  • Hironobu Ihn
    • 1
  1. 1.Department of Dermatology and Plastic Surgery, Faculty of Life SciencesKumamoto UniversityKumamotoJapan

Personalised recommendations