Knockdown of elF3a inhibits TGF‑β1‑induced extracellular matrix protein expression in keloid fibroblasts

Li,Tianyu; Zhao,Junxiang

doi:10.3892/mmr.2017.8365

Knockdown of elF3a inhibits TGF‑β1‑induced extracellular matrix protein expression in keloid fibroblasts

Authors:
- Tianyu Li
- Junxiang Zhao
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Affiliations: Plastic and Cosmetic Center, Nanyang Nanshi Hospital, Affiliated Hospital of Henan University, Nanyang, Henan 473001, P.R. China
Published online on: December 29, 2017 https://doi.org/10.3892/mmr.2017.8365
Pages: 4057-4061

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Abstract

Keloid formation is characterized by hyperproliferation of secretory and responsive keloid fibroblasts (KFs) and overproduction of extracellular matrix (ECM). Eukaryotic translation initiation factor 3 subunit A (eIF3a) one of the core subunits of the translation initiation complex, eIF3, has previously been reported to possess an anti‑fibrogenic effect. However, the role of eIF3a in keloid formation has not yet been investigated. Therefore, the present study examined the effect of eIF3a on transforming growth factor‑β1 (TGF‑β1)‑mediated ECM expression in KFs. The expression levels of eIF3a in human keloid tissues was evaluated using reverse transcription‑quantitative polymerase chain reaction and western blotting. KFs were incubated with siRNA‑eIF3a or siRNA‑mock for 48 h. The cells were then treated with TGF‑β1 (10 ng/ml) for 72 h. Cell proliferation was evaluated using the CCK‑8 assay. The expression levels of α‑SMA, collagen type I, TGF‑β receptor I (RI), TGF‑β RII, phosphorylated (p)‑mothers against decapentaplegic homolog (Smad2), Smad2, p‑Smad3 and Smad3 were detected western blotting. The present study identified significant upregulation of eIF3a mRNA and protein and in human keloid tissues compared with in normal tissues. Knockdown of eIF3a inhibited KF proliferation induced by TGF‑β1. In addition, eIF3a silencing significantly suppressed the TGF‑β1‑induced expression of α‑smooth muscle actin, collagen I, TGF‑β RI and TGF‑β RII in KFs. Furthermore, eIF3a silencing inhibited the phosphorylation levels of Smad2 and Smad3 in TGF‑β1‑induced KFs. To the best of our knowledge, the current study is the first to demonstrate that siRNA‑eIF3a inhibits the expression ECM proteins via the TGF‑β1/Smad signaling pathway in KFs. Therefore, eIF3a may be a potential, novel target for treatment of keloids.

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1	Seifert O and Mrowietz U: Keloid scarring: Bench and bedside. Arch Dermatol Res. 301:259–272. 2009. View Article : Google Scholar : PubMed/NCBI
2	English RS and Shenefelt PD: Keloids and hypertrophic scars. Dermatol Surg. 25:631–638. 1999. View Article : Google Scholar : PubMed/NCBI
3	Gauglitz GG, Korting HC, Pavicic T, Ruzicka T and Jeschke MG: Hypertrophic scarring and keloids: Pathomechanisms and current and emerging treatment strategies. Mol Med. 17:113–125. 2011. View Article : Google Scholar : PubMed/NCBI
4	Fang F, Huang RL, Zheng Y, Liu M and Huo R: Bone marrow derived mesenchymal stem cells inhibit the proliferative and profibrotic phenotype of hypertrophic scar fibroblasts and keloid fibroblasts through paracrine signaling. J Dermatol Sci. 83:95–105. 2016. View Article : Google Scholar : PubMed/NCBI
5	Wu CS, Wu PH, Fang AH and Lan CC: FK506 inhibits the enhancing effects of transforming growth factor (TGF)-β1 on collagen expression and TGF-β/Smad signaling in keloid fibroblasts: Implication for new therapeutic approach. Br J Dermatol. 167:532–541. 2012. View Article : Google Scholar : PubMed/NCBI
6	Makino S, Mitsutake N, Nakashima M, Saenko VA, Ohtsuru A, Umezawa K, Tanaka K, Hirano A and Yamashita S: DHMEQ, a novel NF-kappaB inhibitor, suppresses growth and type I collagen accumulation in keloid fibroblasts. J Dermatol Sci. 51:171–180. 2008. View Article : Google Scholar : PubMed/NCBI
7	Bran GM, Sommer UJ, Goessler UR, Hörmann K, Riedel F and Sadick H: TGF-β1 antisense impacts the SMAD signalling system in fibroblasts from keloid scars. Anticancer Res. 30:3459–3463. 2010.PubMed/NCBI
8	Chalmers RL: The evidence for the role of transforming growth factor-beta in the formation of abnormal scarring. Int Wound J. 8:218–223. 2011. View Article : Google Scholar : PubMed/NCBI
9	Peltonen J, Hsiao LL, Jaakkola S, Sollberg S, Aumailley M, Timpl R, Chu ML and Uitto J: Activation of collagen gene expression in keloids: Co-localization of type I and VI collagen and transforming growth factor-beta 1 mRNA. J Invest Dermatol. 97:240–248. 1991. View Article : Google Scholar : PubMed/NCBI
10	Chin GS, Liu W, Peled Z, Lee TY, Steinbrech DS, Hsu M and Longaker MT: Differential expression of transforming growth factor-beta receptors I and II and activation of Smad 3 in keloid fibroblasts. Plast Reconstr Surg. 108:423–429. 2001. View Article : Google Scholar : PubMed/NCBI
11	Saletta F, Suryo Rahmanto Y and Richardson DR: The translational regulator eIF3a: The tricky eIF3 subunit! Biochim Biophys Acta 1806. 1–286. 2010.
12	Dong Z, Liu Z, Cui P, Pincheira R, Yang Y, Liu J and Zhang JT: Role of eIF3a in regulating cell cycle progression. Exp Cell Res. 315:1889–1894. 2009. View Article : Google Scholar : PubMed/NCBI
13	Liu Z, Dong Z, Yang Z, Chen Q, Pan Y, Yang Y, Cui P, Zhang X and Zhang JT: Role of eIF3a (eIF3 p170) in intestinal cell differentiation and its association with early development. Differentiation. 75:652–661. 2007. View Article : Google Scholar : PubMed/NCBI
14	Zhang YF, Wang Q, Luo J, Yang S, Wang JL and Li HY: Knockdown of elF3a inhibits collagen synthesis in renal fibroblasts via Inhibition of transforming growth factor-β1/Smad signaling pathway. Int Clin Exp Pathol. 8:8983–8989. 2015.
15	Ishihara H, Yoshimoto H, Fujioka M, Murakami R, Hirano A, Fujii T, Ohtsuru A, Namba H and Yamashita S: Keloid fibroblasts resist ceramide-induced apoptosis by overexpression of insulin-like growth factor I receptor. J Invest Dermatol. 115:1065–1071. 2000. View Article : Google Scholar : PubMed/NCBI
16	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
17	Li XW, Wu YH, Li XH, Li D, Du J, Hu CP and Li YJ: Role of eukaryotic translation initiation factor 3a in bleomycin-induced pulmonary fibrosis. Eur J PharSmacol. 749:89–97. 2015. View Article : Google Scholar
18	Li WQ, Li XH, Wu YH, Du J, Wang AP, Li D and Li YJ: Role of eukaryotic translation initiation factors 3a in hypoxia-induced right ventricular remodeling of rats. Life Sci. 144:61–68. 2016. View Article : Google Scholar : PubMed/NCBI
19	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. 33:148–154. 1994. View Article : Google Scholar : PubMed/NCBI
20	Sidgwick G and Bayat A: Extracellular matrix molecules implicated in hypertrophic and keloid scarring. J Eur Acad Dermatol Venereol. 26:141–152. 2012. View Article : Google Scholar : PubMed/NCBI
21	Babu M, Diegelmann R and Oliver N: Keloid fibroblasts exhibit an altered response to TGF-beta. J Invest Dermatol. 99:650–655. 1992. View Article : Google Scholar : PubMed/NCBI
22	Fujiwara M, Muragaki Y and Ooshima A: Keloid-derived fibroblasts show increased secretion of factors involved in collagen turnover and depend on matrix metalloproteinase for migration. Br J Dermatol. 153:295–300. 2005. View Article : Google Scholar : PubMed/NCBI
23	Hsu YC, Chen MJ, Yu YM, Ko SY and Chang CC: Suppression of TGF-β1/SMAD pathway and extracellular matrix production in primary keloid fibroblasts by curcuminoids: Its potential therapeutic use in the chemoprevention of keloid. Arch Dermatol Res. 302:717–724. 2010. View Article : Google Scholar : PubMed/NCBI
24	Mun JH, Kim YM, Kim BS, Kim JH, Kim MB and Ko HC: Simvastatin inhibits transforming growth factor-β1-induced expression of type I collagen, CTGF, and α-SMA in keloid fibroblasts. Wound Repair Regen. 22:125–133. 2014. View Article : Google Scholar : PubMed/NCBI
25	Hou Q, He WJ, Hao HJ, Han QW, Chen L, Dong L, Liu JJ, Li X, Zhang YJ, Ma YZ, et al: The four-herb Chinese medicine ANBP enhances wound healing and inhibits scar formation via bidirectional regulation of transformation growth factor pathway. PLoS One. 9:e1122742014. View Article : Google Scholar : PubMed/NCBI
26	Derynck R and Zhang YE: Smad-dependent and Smad-independent pathways in TGF-beta family signalling. Nature. 425:577–584. 2003. View Article : Google Scholar : PubMed/NCBI
27	Phan TT, Lim IJ, Aalami O, Lorget F, Khoo A, Tan EK, Mukhopadhyay A and Longaker MT: Smad3 signalling plays an important role in keloid pathogenesis via epithelial-mesenchymal interactions. J Pathol. 207:232–242. 2005. View Article : Google Scholar : PubMed/NCBI
28	Wang Z, Gao Z, Shi Y, Sun Y, Lin Z, Jiang H, Hou T, Wang Q, Yuan X, Zhu X, et al: Inhibition of Smad3 expression decreases collagen synthesis in keloid disease fibroblasts. J Plast Reconstr Aesthet Surg. 60:1193–1199. 2007. View Article : Google Scholar : PubMed/NCBI