Open Access

Trichostatin A‑induced miR‑30a‑5p regulates apoptosis and proliferation of keloid fibroblasts via targeting BCL2

  • Authors:
    • Xiaoqing Jian
    • Le Qu
    • Yunlin Wang
    • Qianlei Zou
    • Qing Zhao
    • Shuang Chen
    • Xinghua Gao
    • Hongduo Chen
    • Chundi He
  • View Affiliations

  • Published online on: April 24, 2019     https://doi.org/10.3892/mmr.2019.10185
  • Pages: 5251-5262
  • Copyright: © Jian et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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


Abstract

Keloids are benign fibrous overgrowths that occur as a result of abnormal wound healing following cutaneous injury. MicroRNAs (miRNAs/miRs) are short non‑coding RNAs that serve critical roles in numerous important biological processes, such as cell proliferation, differentiation and apoptosis. However, their role in keloid development remains largely unknown. In the present study, the role of miR‑30a‑5p, a miRNA regulated by Trichostatin A (TSA), in apoptosis within cultured keloid fibroblasts was investigated. An MTT assay was used to detect the proliferation of cultured keloid fibroblasts treated with TSA. Cell apoptosis and cell cycle phases were analyzed using flow cytometry. In addition, an miRNA microarray was performed to compare expression profiles between cultured keloid fibroblasts treated with or without 1,000 nM TSA. Reverse transcription‑quantitative polymerase chain reaction analysis was conducted to estimate miRNA expression levels. The direct target of miR‑30a‑5p was identified using a dual‑luciferase reporter assay. Western blotting was employed to assess protein expression levels in keloid fibroblasts. The results demonstrated that TSA inhibited the proliferation of keloid fibroblasts in a time‑ and dose‑dependent manner. The miRNA microarray revealed alterations in the expression of numerous miRNA sequences in response to TSA when compared with controls. Notably, the expression of miR‑30a‑5p was downregulated in keloid tissues. In addition, overexpression of miR‑30a‑5p induced apoptosis by targeting B‑cell lymphoma 2, which was similar to that observed in response to TSA. These results provide important information regarding a novel miR‑30a‑5p‑mediated signaling pathway induced by TSA treatment, and suggest a potential use for TSA and miR‑30a‑5p as effective therapeutic strategies for keloids.

References

1 

Li Z and Jin Z: Comparative effect and safety of verapamil in keloid and hypertrophic scar treatment: A meta-analysis. Ther Clin Risk Manag. 12:1634–1641. 2016. View Article : Google Scholar

2 

Xue M and Jackson CJ: Extracellular matrix reorganization during wound healing and its impact on abnormal scarring. Adv Wound Care (New Rochelle). 4:119–136. 2015. View Article : Google Scholar : PubMed/NCBI

3 

Wilgus TA and Wulff BC: The importance of mast cells in dermal scarring. Adv Wound Care (New Rochelle). 3:356–365. 2014. View Article : Google Scholar : PubMed/NCBI

4 

Younai S, Nichter LS, Wellisz T, Reinisch J, Nimni ME and Tuan TL: Modulation of collagen synthesis by transforming growth factor-beta in keloid and hypertrophic scar fibroblasts. Ann Plast Surg. 33:148–151. 1994. View Article : Google Scholar : PubMed/NCBI

5 

Suarez E, Syed F, Rasgado TA, Walmsley A, Mandal P and Bayat A: Skin equivalent tensional force alters keloid fibroblast behavior and phenotype. Wound Repair Regen. 22:557–68. 2014. View Article : Google Scholar : PubMed/NCBI

6 

Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004. View Article : Google Scholar : PubMed/NCBI

7 

Sonkoly E, Stahle M and Pivarcsi A: MicroRNAs: Novel regulators in skin inflammation. Clin Exp Dermatol. 33:312–315. 2008. View Article : Google Scholar : PubMed/NCBI

8 

Igoucheva O and Alexeev V: MicroRNA-dependent regulation of cKit in cutaneous melanoma. Biochem Biophys Res Commun. 379:790–794. 2009. View Article : Google Scholar : PubMed/NCBI

9 

Molnar V, Tamasi V, Bakos B, Wiener Z and Falus A: Changes in miRNA expression in solid tumors: An miRNA profiling in melanomas. Semin Cancer Biol. 18:111–122. 2008. View Article : Google Scholar : PubMed/NCBI

10 

Mottamal M, Zheng S, Huang TL and Wang G: Histone deacetylase inhibitors in clinical studies as templates for new anticancer agents. Molecules. 20:3898–3941. 2015. View Article : Google Scholar : PubMed/NCBI

11 

Schneider MR: MicroRNAs as novel players in skin development, homeostasis and disease. Br J Dermatol. 166:22–28. 2012. View Article : Google Scholar : PubMed/NCBI

12 

Zhao LM and Zhang JH: Histone deacetylase inhibitors in tumor immunotherapy. Curr Med Chem. 2017.(Epub ahead of prin). View Article : Google Scholar

13 

Singh A, Patel P, Jageshwar, Patel VK, Jain DK, Kamal M and Rajak H: The safety, efficacy and therapeutic potential of histone deacetylase inhibitors with special reference to panobinostat in gastrointestinal tumors: A Review of Preclinical and Clinical Studies. Curr Cancer Drug Targets. 18:720–736. 2018. View Article : Google Scholar : PubMed/NCBI

14 

Yang L, Qu M, Wang Y, Duan H, Chen P, Wang Y, Shi W, Danielson P and Zhou Q: Trichostatin a inhibits transforming growth Factor-β induced reactive oxygen species accumulation and myofibroblast differentiation via enhanced NF-E2-related factor 2-antioxidant response element signaling. Mol Pharmacol. 83:671–680. 2013. View Article : Google Scholar : PubMed/NCBI

15 

Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001. View Article : Google Scholar : PubMed/NCBI

16 

Rao X, Huang X, Zhou Z and Lin X: An improvement of the 2ˆ(-delta delta CT) method for quantitative real-time polymerase chain reaction data analysis. Biostat Bioinforma Biomath. 3:71–85. 2013.PubMed/NCBI

17 

Rhodes LV, Nitschke AM, Segar HC, Martin EC, Driver JL, Elliott S, Nam SY, Li M, Nephew KP, Burow ME and Collins-Burow BM: The histone deacetylase inhibitor trichostatin A alters microRNA expression profiles in apoptosis-resistant breast cancer cells. Oncol Rep. 27:10–16. 2012.PubMed/NCBI

18 

Williams SM, Golden-Mason L, Ferguson BS, Schuetze KB, Cavasin MA, Demos-Davies K, Yeager ME, Stenmark KR and McKinsey TA: Class I HDACs regulate angiotensin II-dependent cardiac fibrosis via fibroblasts and circulating fibrocytes. J Mol Cell Cardiol. 67:112–125. 2014. View Article : Google Scholar : PubMed/NCBI

19 

Bodas M, Mazur S, Min T and Vij N: Inhibition of histone-deacetylase activity rescues inflammatory cystic fibrosis lung disease by modulating innate and adaptive immune responses. Respir Res. 19:22018. View Article : Google Scholar : PubMed/NCBI

20 

Xu X, Jin S, Ma Y, Fan Z, Yan Z, Li W, Song Q, You W, Lyu Z and Song Y: miR-30a-5p enhances paclitaxel sensitivity in non-small cell lung cancer through targeting BCL-2 expression. J Mol Med (Berl). 95:861–871. 2017. View Article : Google Scholar : PubMed/NCBI

21 

Tung CW, Hsu YC, Cai CJ, Shih YH, Wang CJ, Chang PJ and Lin CL: Trichostatin A ameliorates renal tubulointerstitial fibrosis through modulation of the JNK-dependent Notch-2 signaling pathway. Sci Rep. 7:144952017. View Article : Google Scholar : PubMed/NCBI

22 

Kaimori A, Potter JJ, Choti M, Ding Z, Mezey E and Koteish AA: Histone deacetylase inhibition suppresses the transforming growth factor beta1-induced epithelial-to-mesenchymal transition in hepatocytes. Hepatology. 52:1033–45. 2010. View Article : Google Scholar : PubMed/NCBI

23 

Rombouts K, Niki T, Greenwel P, Vandermonde A, Wielant A, Hellemans K, De Bleser P, Yoshida M, Schuppan D, Rojkind M and Geerts A: Trichostatin A, a histone deacetylase inhibitor, suppresses collagen synthesis and prevents TGF-beta(1)-induced fibrogenesis in skin fibroblasts. Exp Cell Res. 278:184–197. 2002. View Article : Google Scholar : PubMed/NCBI

24 

Sun J, Wang Y, Cui W, Lou Y, Sun G, Zhang D and Miao L: Role of epigenetic histone modifications in diabetic kidney disease involving renal fibrosis. J Diabetes Res. 7:242–384. 2017.

25 

Ghosh AK, Mori Y, Dowling E and Varga J: Trichostatin A blocks TGF-beta-induced collagen gene expression in skin fibroblasts: involvement of Sp1. Biochem Biophys Res Commun. 354:420–426. 2007. View Article : Google Scholar : PubMed/NCBI

26 

Diao JS, Xia WS, Yi CG, Wang YM, Li B, Xia W, Liu B, Guo SZ and Sun XD: Trichostatin A inhibits collagen synthesis and induces apoptosis in keloid fibroblasts. Arch Dermatol Res. 303:573–580. 2011. View Article : Google Scholar : PubMed/NCBI

27 

Babalola O, Mamalis A, Lev-Tov H and Jagdeo J: The role of microRNAs in skin fibrosis. Arch Dermatol Res. 305:763–76. 2013. View Article : Google Scholar : PubMed/NCBI

28 

Zhu H, Han C and Wu T: MiR-17-92 cluster promotes hepatocarcinogenesis. Carcinogenesis. 36:1213–22. 2015. View Article : Google Scholar : PubMed/NCBI

29 

Dakhlallah D, Batte K, Wang Y, Cantemir-Stone CZ, Yan P, Nuovo G, Mikhail A, Hitchcock CL, Wright VP, Piper MG and Marsh CB: Epigenetic regulation of miR-17~92 contributes to the pathogenesis of pulmonary fibrosis. Am J Respir Crit Care Med. 187:397–405. 2013. View Article : Google Scholar : PubMed/NCBI

30 

Li YL, Wang J, Zhang CY, Shen YQ, Wang HM, Ding L, Gu YC, Lou JT, Zhao XT, Ma ZL and Jin YX: MiR-146a-5p inhibits cell proliferation and cell cycle progression in NSCLC cell lines by targeting CCND1 and CCND2. Eur Respir J. 7:59287–59298. 2016.

31 

He R, Yang L, Lin X, Chen X, Lin X, Wei F, Liang X, Luo Y, Wu Y, Gan T, et al: MiR-30a-5p suppresses cell growth and enhances apoptosis of hepatocellular carcinoma cells via targeting AEG-1. Int J Clin Exp Pathol. 8:15632–15641. 2015.PubMed/NCBI

32 

Wang Z, Dai X, Chen Y, Sun C, Zhu Q, Zhao H, Liu G, Huang Q and Lan Q: MiR-30a-5p is induced by Wnt/β-catenin pathway and promotes glioma cell invasion by repressing NCAM. Biochem Biophys Res Commun. 465:374–380. 2015. View Article : Google Scholar : PubMed/NCBI

33 

Safaeian L, Abed A and Vaseghi G: The role of Bcl-2 family proteins in pulmonary fibrosis. Eur J Pharmacol. 741:281–9. 2014. View Article : Google Scholar : PubMed/NCBI

Related Articles

Journal Cover

June 2019
Volume 19 Issue 6

Print ISSN: 1791-2997
Online ISSN:1791-3004

Sign up for eToc alerts

Recommend to Library

Copy and paste a formatted citation
APA
Jian, X., Qu, L., Wang, Y., Zou, Q., Zhao, Q., Chen, S. ... He, C. (2019). Trichostatin A‑induced miR‑30a‑5p regulates apoptosis and proliferation of keloid fibroblasts via targeting BCL2. Molecular Medicine Reports, 19, 5251-5262. https://doi.org/10.3892/mmr.2019.10185
MLA
Jian, X., Qu, L., Wang, Y., Zou, Q., Zhao, Q., Chen, S., Gao, X., Chen, H., He, C."Trichostatin A‑induced miR‑30a‑5p regulates apoptosis and proliferation of keloid fibroblasts via targeting BCL2". Molecular Medicine Reports 19.6 (2019): 5251-5262.
Chicago
Jian, X., Qu, L., Wang, Y., Zou, Q., Zhao, Q., Chen, S., Gao, X., Chen, H., He, C."Trichostatin A‑induced miR‑30a‑5p regulates apoptosis and proliferation of keloid fibroblasts via targeting BCL2". Molecular Medicine Reports 19, no. 6 (2019): 5251-5262. https://doi.org/10.3892/mmr.2019.10185