Downregulation of activin‑signaling gene expression in passaged normal human dermal fibroblasts

Kim,Young  Il; Lee,Chan‑Yang; Shin,Min  Kyung

doi:10.3892/br.2019.1258

Downregulation of activin‑signaling gene expression in passaged normal human dermal fibroblasts

Authors:
- Young Il Kim
- Chan‑Yang Lee
- Min Kyung Shin
View Affiliations

Affiliations: Medical Science Research Institute, Kyung Hee University Medical Center, Seoul 02447, Republic of Korea, Department of Dermatology, College of Medicine, Kyung Hee University, Seoul 02447, Republic of Korea
Published online on: November 21, 2019 https://doi.org/10.3892/br.2019.1258
Pages: 17-22

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

Activins are members of the transforming growth factor‑β (TGF‑β) superfamily and play important roles in proliferation, differentiation, and apoptosis of various target cells. We investigated changes of activin, activin receptor (ActR), and Smad‑signaling gene expression with increasing passage number in normal human dermal fibroblasts. The expression of mRNA and protein was measured by reverse transcription‑quantitative polymerase chain reaction and immunoblot analysis from passage numbers 5 to 15. Activin A and follistatin transcript levels increased with increasing passage number. ActR types IA, IB, IIA and IIB mRNA levels decreased at high passage number. The levels of Smad2, 3 and 4 protein decreased with increasing passage number, which also attenuated phosphorylation of Smad2 and 3 protein expression. Smad7 was enhanced with increasing passage number. These results suggest that expression of activin‑signaling in aging normal human dermal fibroblasts increases activin A and follistatin, whereas ActR‑Smad signaling is decreased.

View Figures

View References

1	DePaolo LV, Bicsak TA, Erickson GF, Shimasaki S and Ling N: Follistatin and activin: A potential intrinsic regulatory system within diverse tissues. Proc Soc Exp Biol Med. 198:500–512. 1991.PubMed/NCBI View Article : Google Scholar
2	Xia Y and Schneyer AL: The biology of activin: Recent advances in structure, regulation and function. J Endocrinol. 202:1–12. 2009.PubMed/NCBI View Article : Google Scholar
3	McDowall M, Edwards NM, Jahoda CA and Hynd PI: The role of activins and follistatins in skin and hair follicle development and function. Cytokine Growth Factor Rev. 19:415–426. 2008.PubMed/NCBI View Article : Google Scholar
4	Kawakami S, Fujii Y and Winters SJ: Follistatin production by skin fibroblasts and its regulation by dexamethasone. Mol Cell Endocrinol. 172:157–167. 2001.PubMed/NCBI View Article : Google Scholar
5	Attisano L, Wrana JL, Montalvo E and Massagué J: Activation of signalling by the activin receptor complex. Mol Cell Biol. 16:1066–1073. 1996.PubMed/NCBI View Article : Google Scholar
6	Moustakas A, Souchelnytskyi S and Heldin CH: Smad regulation in TGF-β signal transduction. J Cell Sci. 114:4359–4369. 2001.PubMed/NCBI
7	Nakao A, Afrakhte M, Morén A, Nakayama T, Christian JL, Heuchel R, Itoh S, Kawabata M, Heldin NE, Heldin CH, et al: Identification of Smad7, a TGFbeta-inducible antagonist of TGF-beta signalling. Nature. 389:631–635. 1997.PubMed/NCBI View Article : Google Scholar
8	Afrakhte M, Morén A, Jossan S, Itoh S, Sampath K, Westermark B, Heldin CH, Heldin NE and ten Dijke P: Induction of inhibitory Smad6 and Smad7 mRNA by TGF-β family members. Biochem Biophys Res Commun. 249:505–511. 1998.PubMed/NCBI View Article : Google Scholar
9	Kim KS, Park HK, Lee JW, Kim YI and Shin MK: Investigate correlation between mechanical property and aging biomarker in passaged human dermal fibroblasts. Microsc Res Tech. 78:277–282. 2015.PubMed/NCBI View Article : Google Scholar
10	Kuchler RJ: Development of animal cell populations in vitro. In: Biochemical Methods in Cell Culture and Virology. Kuchler RJ (ed). Dowden, Hutchinson & Ross, Inc., Stroudsburg PA, pp90-113. 1977.
11	Sjerobabski-Masnec I and Situm M: Skin aging. Acta Clin Croat. 49:515–518. 2010.PubMed/NCBI
12	Moura J, da Silva L, Cruz MT and Carvalho E: Molecular and cellular mechanisms of bone morphogenetic proteins and activins in the skin: Potential benefits for wound healing. Arch Dermatol Res. 305:557–569. 2013.PubMed/NCBI View Article : Google Scholar
13	Beer HD, Gassmann MG, Munz B, Steiling H, Engelhardt F, Bleuel K and Werner S: Expression and function of keratinocyte growth factor and activin in skin morphogenesis and cutaneous wound repair. J Investig Dermatol Symp Proc. 5:34–39. 2000.PubMed/NCBI View Article : Google Scholar
14	Antsiferova M, Huber M, Meyer M, Piwko-Czuchra A, Ramadan T, MacLeod AS, Havran WL, Dummer R, Hohl D and Werner S: Activin enhances skin tumourigenesis and malignant progression by inducing a pro-tumourigenic immune cell response. Nat Commun. 2(576)2011.PubMed/NCBI View Article : Google Scholar
15	Kojima I, Maeshima A and Zhang YQ: Role of the activin-follistatin system in the morphogenesis and regeneration of the renal tubules. Mol Cell Endocrinol. 180:179–182. 2001.PubMed/NCBI View Article : Google Scholar
16	Refaat B, Ashshi AM, El-Shemi AG and Azhar E: Activins and Follistatin in Chronic Hepatitis C and Its Treatment with Pegylated-Interferon-α Based Therapy. Mediators Inflamm. 2015(287640)2015.PubMed/NCBI View Article : Google Scholar
17	Santoro N, Adel T and Skurnick JH: Decreased inhibin tone and increased activin A secretion characterize reproductive aging in women. Fertil Steril. 71:658–662. 1999.PubMed/NCBI View Article : Google Scholar
18	Barrios-Silva LV, Parnell M, Shinwari ZB, Chaudhary GA, Xenofontos T, van Bekhoven A, McArthur S and Elliott BT: Activin subfamily peptides predict chronological age in humans. Physiol Rep. 6(e13823)2018.PubMed/NCBI View Article : Google Scholar
19	Hedger MP and de Kretser DM: The activins and their binding protein, follistatin-Diagnostic and therapeutic targets in inflammatory disease and fibrosis. Cytokine Growth Factor Rev. 24:285–295. 2013.PubMed/NCBI View Article : Google Scholar
20	Massagué J: TGF-β signal transduction. Annu Rev Biochem. 67:753–791. 1998.PubMed/NCBI View Article : Google Scholar
21	Fisher GJ, Shao Y, He T, Qin Z, Perry D, Voorhees JJ and Quan T: Reduction of fibroblast size/mechanical force down-regulates TGF-β type II receptor: Implications for human skin aging. Aging Cell. 15:67–76. 2016.PubMed/NCBI View Article : Google Scholar
22	Zhao H, Patra A, Tanaka Y, Li LC and Dahiya R: Transforming growth factor-β(s) and their receptors in aging rat prostate. Biochem Biophys Res Commun. 294:464–469. 2002.PubMed/NCBI View Article : Google Scholar
23	Han KH, Choi HR, Won CH, Chung JH, Cho KH, Eun HC and Kim KH: Alteration of the TGF-β/SMAD pathway in intrinsically and UV-induced skin aging. Mech Ageing Dev. 126:560–567. 2005.PubMed/NCBI View Article : Google Scholar
24	Hübner G, Hu Q, Smola H and Werner S: Strong induction of activin expression after injury suggests an important role of activin in wound repair. Dev Biol. 173:490–498. 1996.PubMed/NCBI View Article : Google Scholar
25	Hata A and Chen YG: TGF-β Signaling from Receptors to Smads. Cold Spring Harb Perspect Biol. 8(a022061)2016.PubMed/NCBI View Article : Google Scholar
26	Massagué J, Seoane J and Wotton D: Smad transcription factors. Genes Dev. 19:2783–2810. 2005.PubMed/NCBI View Article : Google Scholar
27	Matsuo SE, Fiore AP, Siguematu SM, Ebina KN, Friguglietti CU, Ferro MC, Kulcsar MA and Kimura ET: Expression of SMAD proteins, TGF-beta/activin signaling mediators, in human thyroid tissues. Arq Bras Endocrinol Metabol. 54:406–412. 2010.PubMed/NCBI View Article : Google Scholar
28	Choi MJ, Song KM, Park JM, Kwon MH, Kwon KD, Park SH, Ryu DS, Ryu JK and Suh JK: Effect of SMAD7 gene overexpression on TGF-β1-induced profibrotic responses in fibroblasts derived from Peyronie's plaque. Asian J Androl. 17:487–492. 2015.PubMed/NCBI View Article : Google Scholar
29	Kim YI, Kim KS, Ahn HJ, Kang IH and Shin MK: Reduced matrix metalloproteinase and collagen transcription mediated by the TGF-β/Smad pathway in passaged normal human dermal fibroblasts. J Cosmet Dermatol. Sep 11. 2019.(Epub ahead of print). PubMed/NCBI View Article : Google Scholar