Downregulation of miR-542-3p Contributes to Apoptosis Resistance in Dermal Fibroblasts from Systemic Sclerosis Patients via Survivin Overexpression

Keywords: Apoptosis, miR-542-3p, Survivin, Systemic sclerosis


Systemic sclerosis (SSc) is characterized by excessive production of collagens by fibroblasts that leads to vast fibrosis. Resistance to apoptosis is one of the possible underlying mechanisms of fibrosis in these patients. Survivinis involved in inhibition of apoptosis and aberrantly functions in SSc. Since dysregulation of survivin-targeting microRNAs (miRNAs) has frequently been observed in cancer and some autoimmune disorders, this study aimed to investigate their expression status in dermal fibroblasts from SSc patients. DiffuseSSc patients were selected according to American College of Rheumatology criteria. Isolated fibroblasts from 10 SSc and 10 healthy skin biopsies were cultured. After examining purity of the cells, mRNA and miRNAs extraction was performed followed by complementary DNA (cDNA) synthesis. Relative expressions ofsurvivin mRNA, miR-16-5p, miR-320a, miR-218-5p, miR-708-5p and miR-542-3p were analyzed using real time PCR. Survivin mRNA expression was significantly 1.85-fold upregulated in fibroblasts from SSc patients compared with healthy controls (p=0.046). Among the studied miRNAs, miR-542-3p expression was significantly decreased (p=0.033), while enhanced expression of miR-708-5p was observed in SSc fibroblasts (p=0.05) in comparison to healthy subjects. Downregulation of miR-542-3p significantly correlated with survivin overexpression (r=˗0.45, p=0.049). Downregulation of miR-542-3p that is correlated with higher surviving expression levels might be a possible cause of apoptosis resistance in SSc fibroblasts, hence providing a new understanding of the disease pathogenesis.


1. LeRoy EC, Black C, Fleischmajer R, Jablonska S, Krieg T, Medsger TA, Jr., et al. Scleroderma (systemic sclerosis): classification, subsets and pathogenesis. The Journal of rheumatology. 1988;15(2):202-5.

2. Zuo XX, Zhang LH, Luo H, Li Y, Zhu H. Systematic approach to understanding the pathogenesis of systemic sclerosis. 2016.

3. Mahmoudi M, Fallahian F, Sobhani S, Ghoroghi S, Jamshidi A, Poursani S, et al. Analysis of killer cell immunoglobulin-like receptors (KIRs) and their HLA ligand genes polymorphisms in Iranian patients with systemic sclerosis. Clinical rheumatology. 2017;36(4):853-62.

4. Aslani S, Mahmoudi M, Karami J, Jamshidi AR, Malekshahi Z, Nicknam MH. Epigenetic alterations underlying autoimmune diseases. Autoimmunity. 2016;49(2):69-83.

5. Karimizadeh E, Gharibdoost F, Motamed N, Jafarinejad-Farsangi S, Jamshidi A, Mahmoudi M. c-Abl silencing reduced the inhibitory effects of TGF-β1 on apoptosis in systemic sclerosis dermal fibroblasts. Molecular and cellular biochemistry. 2015;405(1-2):169-76.

6. Karimizadeh E, Motamed N, Mahmoudi M, Jafarinejad-Farsangi S, Jamshidi A, Faridani H, et al. Attenuation of fibrosis with selective inhibition of c-Abl by siRNA in systemic sclerosis dermal fibroblasts. Archives of dermatological research. 2015;307(2):135-42.

7. Darby IA, Hewitson TD. Fibroblast differentiation in wound healing and fibrosis. International review of cytology. 2007;257:143-79.

8. Jelaska A, Korn JH. Role of apoptosis and transforming growth factor beta1 in fibroblast selection and activation in systemic sclerosis. Arthritis and rheumatism. 2000;43(10):2230-9.

9. Chabaud S, Moulin VJ. Apoptosis modulation as a promising target for treatment of systemic sclerosis. International journal of rheumatology. 2011;2011:495792.

10. Jun JB, Kuechle M, Harlan JM, Elkon KB. Fibroblast and endothelial apoptosis in systemic sclerosis. Current opinion in rheumatology. 2003;15(6):756-60.

11. Kissin E, Korn JH. Apoptosis and myofibroblasts in the pathogenesis of systemic sclerosis. Current rheumatology reports. 2002;4(2):129-35.

12. Wheatley SP, McNeish IA. Survivin: a protein with dual roles in mitosis and apoptosis. International review of cytology. 2005;247:35-88.

13. Altieri DC. Survivin - The inconvenient IAP. Seminars in cell & developmental biology. 2015;39:91-6.

14. Boidot R, Vegran F, Lizard-Nacol S. Transcriptional regulation of the survivin gene. Molecular biology reports. 2014;41(1):233-40.

15. Sah NK, Khan Z, Khan GJ, Bisen PS. Structural, functional and therapeutic biology of survivin. Cancer letters. 2006;244(2):164-71.

16. Srinivasula SM, Ashwell JD. IAPs: what's in a name? Molecular cell. 2008;30(2):123-35.

17. Zangemeister-Wittke U, Simon HU. An IAP in action: the multiple roles of survivin in differentiation, immunity and malignancy. Cell cycle (Georgetown, Tex). 2004;3(9):1121-3.

18. Mokuda S, Miyazaki T, Ubara Y, Kanno M, Sugiyama E, Takasugi K, et al. CD1a+ survivin+ dendritic cell infiltration in dermal lesions of systemic sclerosis. Arthritis research & therapy. 2015;17:275.

19. Bartel DP. MicroRNAs: target recognition and regulatory functions. Cell. 2009;136(2):215-33.

20. Mahmoudi M, Aslani S, Nicknam MH, Karami J, Jamshidi AR. New insights toward the pathogenesis of ankylosing spondylitis; genetic variations and epigenetic modifications. Modern rheumatology. 2017;27(2):198-209.

21. Ceribelli A, Satoh M, Chan EK. MicroRNAs and autoimmunity. Current opinion in immunology. 2012;24(6):686-91.

22. Zhang M, Yang J, Li F. Transcriptional and post-transcriptional controls of survivin in cancer cells: novel approaches for cancer treatment. Journal of experimental & clinical cancer research : CR. 2006;25(3):391-402.

23. Huang J, Lyu H, Wang J, Liu B. MicroRNA regulation and therapeutic targeting of survivin in cancer. American journal of cancer research. 2015;5(1):20-31.

24. Zarogoulidis P, Petanidis S, Kioseoglou E, Domvri K, Anestakis D, Zarogoulidis K. MiR-205 and miR-218 expression is associated with carboplatin chemoresistance and regulation of apoptosis via Mcl-1 and Survivin in lung cancer cells. Cellular Signalling. 2015;27(8):1576-88.

25. Hu Y, Xu K, Yague E. miR-218 targets survivin and regulates resistance to chemotherapeutics in breast cancer. Breast cancer research and treatment. 2015;151(2):269-80.

26. Alajez NM, Lenarduzzi M, Ito E, Hui AB, Shi W, Bruce J, et al. MiR-218 suppresses nasopharyngeal cancer progression through downregulation of survivin and the SLIT2-ROBO1 pathway. Cancer research. 2011;71(6):2381-91.

27. Sun JY, Huang Y, Li JP, Zhang X, Wang L, Meng YL, et al. MicroRNA-320a suppresses human colon cancer cell proliferation by directly targeting beta-catenin. Biochemical and biophysical research communications. 2012;420(4):787-92.

28. Lafyatis R. Connective tissue disease: SSc-fibrosis takes flight with Wingless inhibition. Nature reviews Rheumatology. 2012;8(8):441-2.

29. Liu Q FH, Sun F, Zhang H, Tie Y, Zhu J, et al. miR-16 family induces cell cycle arrest by regulating multiple cell cycle genes. Nucleic acids research. 2008;36(16):5391-404.

30. Saini S, Yamamura S, Majid S, Shahryari V, Hirata H, Tanaka Y, et al. MicroRNA-708 induces apoptosis and suppresses tumorigenicity in renal cancer cells. Cancer research. 2011;71(19):6208-19.

31. Griffith TS, Stokes B, Kucaba TA, Earel JK, Jr., VanOosten RL, Brincks EL, et al. TRAIL gene therapy: from preclinical development to clinical application. Current gene therapy. 2009;9(1):9-19.

32. Yoon S CY, Lee S, Jeong Y, Yoon J, Baek K. Induction of growth arrest by miR-542-3p that targets survivin. FEBS Lett. 2010;584(18):4048-52.

33. Preliminary criteria for the classification of systemic sclerosis (scleroderma). Subcommittee for scleroderma criteria of the American Rheumatism Association Diagnostic and Therapeutic Criteria Committee. Arthritis and rheumatism. 1980;23(5):581-90.

34. Schmittgen TDaKJL. Analyzing real-time PCR data by the comparative CT method. Nature protocols. 2008;3(6):1101-8.

35. Santiago B GM, Rivero M, Pablos JL. Decreased susceptibility to Fas-induced apoptosis of systemic sclerosis dermal fibroblasts. Arthritis and rheumatism. 2001;44(7):1667-76.

36. Chen J, Liu M, Mao N, Zeng Y. [Expression of survivin in rheumatoid arthritis]. Zhong nan da xue xue bao Yi xue ban = Journal of Central South University Medical sciences. 2013;38(2):186-90.

37. Isgren A, Forslind K, Erlandsson M, Axelsson C, Andersson S, Lund A, et al. High survivin levels predict poor clinical response to infliximab treatment in patients with rheumatoid arthritis. Seminars in arthritis and rheumatism. 2012;41(5):652-7.

38. Jafarinejad-Farsangi S, Farazmand A, Mahmoudi M, Gharibdoost F, Karimizadeh E, Noorbakhsh F, et al. MicroRNA-29a induces apoptosis via increasing the Bax:Bcl-2 ratio in dermal fibroblasts of patients with systemic sclerosis. Autoimmunity. 2015;48(6):369-78.

39. Jafarinejad-Farsangi S, Farazmand A, Gharibdoost F, Karimizadeh E, Noorbakhsh F, Faridani H, et al. Inhibition of MicroRNA-21 induces apoptosis in dermal fibroblasts of patients with systemic sclerosis. International journal of dermatology. 2016;55(11):1259-67.

40. Ma Q, Wang X, Li Z, Li B, Ma F, Peng L, et al. microRNA-16 represses colorectal cancer cell growth in vitro by regulating the p53/survivin signaling pathway. Oncology reports. 2013;29(4):1652-8.

41. Yoon S, Choi YC, Lee S, Jeong Y, Yoon J, Baek K. Induction of growth arrest by miR-542-3p that targets survivin. FEBS Lett. 2010;584(18):4048-52.

42. Althoff K, Lindner S, Odersky A, Mestdagh P, Beckers A, Karczewski S, et al. miR-542-3p exerts tumor suppressive functions in neuroblastoma by downregulating Survivin. International journal of cancer. 2015;136(6):1308-20.

43. Kureel J, Dixit M, Tyagi AM, Mansoori MN, Srivastava K, Raghuvanshi A, et al. miR-542-3p suppresses osteoblast cell proliferation and differentiation, targets BMP-7 signaling and inhibits bone formation. Cell death & disease. 2014;5:e1050.

44. Landgraf P, Rusu M, Sheridan R, Sewer A, Iovino N, Aravin A, et al. A mammalian microRNA expression atlas based on small RNA library sequencing. Cell. 2007;129(7):1401-14.

45. Schotte D, Chau J, Sylvester G, Liu G, Chen C, Van Der Velden V, et al. Identification of new microRNA genes and aberrant microRNA profiles in childhood acute lymphoblastic leukemia. Leukemia. 2009;23(2):313.

46. Patnaik SK, Kannisto E, Knudsen S, Yendamuri S. Evaluation of microRNA expression profiles that may predict recurrence of localized stage I non–small cell lung cancer after surgical resection. Cancer research. 2010;70(1):36-45.

How to Cite
Vahidi Manesh P, Farazmand A, Gharibdoost F, Vanaki N, Mostafaei S, Kavosi H, Mahmoudi MB, Mahmoudi M. Downregulation of miR-542-3p Contributes to Apoptosis Resistance in Dermal Fibroblasts from Systemic Sclerosis Patients via Survivin Overexpression. Iran J Allergy Asthma Immunol. 18(2):173-181.
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