Suppression of cell proliferation and collagen production in cultured human hypertrophic scar fibroblasts by Sp1 decoy oligodeoxynucleotide

Deng,Chenliang; Zheng,Jianghong; Wan,Weidong; Zhang,Shixin; Ding,Zhi; Mao,Guangyu; Yang,Songlin

doi:10.3892/mmr.2013.1278

Suppression of cell proliferation and collagen production in cultured human hypertrophic scar fibroblasts by Sp1 decoy oligodeoxynucleotide

Authors:
- Chenliang Deng
- Jianghong Zheng
- Weidong Wan
- Shixin Zhang
- Zhi Ding
- Guangyu Mao
- Songlin Yang
View Affiliations

Affiliations: Department of Plastic Surgery, Shanghai 6th People's Hospital, Shanghai Jiaotong University, Shanghai 200233, P.R. China
Published online on: January 18, 2013 https://doi.org/10.3892/mmr.2013.1278
Pages: 785-790

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

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

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Hypertrophic scars are characterized by the abnormal proliferation of fibroblasts and an overproduction of collagen. The Sp1 transcription factor is involved in the stimulation of collagen synthesis. A decoy oligonucleotide (ODN) targeting Sp1 was designed and transfected into hypertrophic scar fibroblasts (HSFs) cells using cationic liposomes. The transfection efficiency was determined by flow cytometry and was observed to be 85±7% (n=5). Specific binding of the Sp1 decoy ODN was monitored with an electrophoretic mobility shift assay (EMSA). Following transfection with the decoy ODN to Sp1, cell viability and cell proliferation, which were examined by the cell counting kit WST‑8, were decreased by 80% compared with untreated cells. Transforming growth factor‑β (TGF‑β) mRNA and collagen mRNA expression were also reduced by 48% in the transfection decoy ODN group. The cell viability of HSFs after 48 h of transfection with 25, 50, 100 and 150 nM Sp1 decoy ODN was 0.9331±0.0203, 0.7479±0.0868, 0.577±0.0347 and 0.4703±0.0147, respectively. The 100 nM dose of the Sp1 decoy ODN inhibited the expression of types I and III collagen by 32 and 28%, respectively (both P<0.01). TGF‑β mRNA expression was also effectively suppressed by the 100 nM Sp1 decoy ODN (P<0.01). The Sp1 decoy ODN inhibited cell proliferation and the expression of types I and III collagen. Therefore, Sp1 decoy ODNs may be a promising tool for developing and testing novel therapeutic applications for treating hypertrophic scars.

View Figures

View References

1	Wang R, Ghahary A, Shen Q, Scott PG, Roy K and Tredget EE: Hypertrophic scar tissues and fibroblasts produce more transforming growth factor-beta1 mRNA and protein than normal skin and cells. Wound Repair Regen. 8:128–137. 2000. View Article : Google Scholar : PubMed/NCBI
2	Jagadeesan J and Bayat A: Transforming growth factor beta (TGFbeta) and keloid disease. Int J Surg. 5:278–285. 2007. View Article : Google Scholar : PubMed/NCBI
3	Chin GS, Liu W, Peled Z, et al: 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
4	Chen SJ, Artlett CM, Jimenez SA and Varga J: Modulation of human alpha1(I) procollagen gene activity by interaction with Sp1 and Sp3 transcription factors in vitro. Gene. 215:101–110. 1998. View Article : Google Scholar : PubMed/NCBI
5	Boros DL, Singh KP, Gerard HC, Hudson AP, White SL and Cutroneo KR: A novel nonsteroidal antifibrotic oligo decoy containing the TGF-beta element found in the COL1A1 gene which regulates murine schistosomiasis liver fibrosis. J Cell Physiol. 204:370–374. 2005. View Article : Google Scholar
6	Chen SJ, Yuan W, Lo S, Trojanowska M and Varga J: Interaction of smad3 with a proximal smad-binding element of the human alpha2(I) procollagen gene promoter required for transcriptional activation by TGF-beta. J Cell Physiol. 183:381–392. 2000. View Article : Google Scholar : PubMed/NCBI
7	Naim R, Naumann A, Barnes J, et al: Transforming growth factor-beta1-antisense modulates the expression of hepatocyte growth factor/scatter factor in keloid fibroblast cell culture. Aesthetic Plast Surg. 32:346–352. 2008. View Article : Google Scholar : PubMed/NCBI
8	Meisler NT, Chiu JF and Cutroneo KR: Promoter competitors as novel antifibrotics that inhibit transforming growth factor-beta induction of collagen and noncollagen protein synthesis in fibroblasts. J Cell Biochem. 75:196–205. 1999. View Article : Google Scholar : PubMed/NCBI
9	Bowley E, O’Gorman DB and Gan BS: Beta-catenin signaling in fibroproliferative disease. J Surg Res. 138:141–150. 2007. View Article : Google Scholar : PubMed/NCBI
10	Smith RA, Miller TM, Yamanaka K, et al: Antisense oligonucleotide therapy for neurodegenerative disease. J Clin Invest. 116:2290–2296. 2006. View Article : Google Scholar : PubMed/NCBI
11	Lindow M and Kauppinen S: Discovering the first microRNA-targeted drug. J Cell Biol. 199:407–412. 2012.PubMed/NCBI
12	Sato M: Upregulation of the Wnt/beta-catenin pathway induced by transforming growth factor-beta in hypertrophic scars and keloids. Acta Derm Venereol. 86:300–307. 2006. View Article : Google Scholar : PubMed/NCBI
13	Takamizawa J, Konishi H, Yanagisawa K, et al: Reduced expression of the let-7 microRNAs in human lung cancers in association with shortened postoperative survival. Cancer Res. 64:3753–3756. 2004. View Article : Google Scholar : PubMed/NCBI
14	Calin GA, Dumitru CD, Shimizu M, et al: Frequent deletions and down-regulation of micro- RNA genes miR15 and miR16 at 13q14 in chronic lymphocytic leukemia. Proc Natl Acad Sci USA. 99:15524–15529. 2002. View Article : Google Scholar : PubMed/NCBI
15	Ahn JD, Kim CH, Magae J, et al: E2F decoy oligodeoxynucleotides effectively inhibit growth of human tumor cells. Biochem Biophys Res Commun. 310:1048–1053. 2003. View Article : Google Scholar : PubMed/NCBI
16	Ahn JD, Morishita R, Kaneda Y, et al: Inhibitory effects of novel AP-1 decoy oligodeoxynucleotides on vascular smooth muscle cell proliferation in vitro and neointimal formation in vivo. Circ Res. 90:1325–1332. 2002. View Article : Google Scholar : PubMed/NCBI
17	Nusse R: Wnt signaling in disease and in development. Cell Res. 15:28–32. 2005. View Article : Google Scholar : PubMed/NCBI
18	Kato M, Zhang J, Wang M, et al: MicroRNA-192 in diabetic kidney glomeruli and its function in TGF-beta-induced collagen expression via inhibition of E-box repressors. Proc Natl Acad Sci USA. 104:3432–3437. 2007. View Article : Google Scholar : PubMed/NCBI
19	Cheon SS, Wei Q, Gurung A, et al: Beta-catenin regulates wound size and mediates the effect of TGF-beta in cutaneous healing. FASEB J. 20:692–701. 2006. View Article : Google Scholar : PubMed/NCBI
20	Berezikov E, Guryev V, van de Belt J, Wienholds E, Plasterk RH and Cuppen E: Phylogenetic shadowing and computational identification of human microRNA genes. Cell. 120:21–24. 2005. View Article : Google Scholar : PubMed/NCBI