Lung fibroblasts from patients with emphysema show markers of senescence in vitro
Tóm tắt
The loss of alveolar walls is a hallmark of emphysema. As fibroblasts play an important role in the maintenance of alveolar structure, a change in fibroblast phenotype could be involved in the pathogenesis of this disease. In a previous study we found a reduced
Primary lung fibroblasts were obtained from 13 patients with moderate to severe lung emphysema (E) and 15 controls (C) undergoing surgery for lung tumor resection or volume reduction (n = 2). Fibroblasts (8E/9C) were stained for senescence-associated β-galactosidase (SA-β-Gal). In independent cultures, DNA from lung fibroblasts (7E/8C) was assessed for mean telomere length. Two exploratory 12 k cDNA microarrays were used to assess gene expression in pooled fibroblasts (3E/3C). Subsequently, expression of selected genes was evaluated by quantitative PCR (qPCR) in fibroblasts of individual patients (10E/9C) and protein concentration was analyzed in the cell culture supernatant.
The median (quartiles) percentage of fibroblasts positive for SA-β-Gal was 4.4 (3.2;4.7) % in controls and 16.0 (10.0;24.8) % in emphysema (p = 0.001), while telomere length was not different. Among the candidates for differentially expressed genes in the array (factor ≥ 3), 15 were upregulated and 121 downregulated in emphysema. qPCR confirmed the upregulation of insulin-like growth factor-binding protein (IGFBP)-3 and IGFBP-rP1 (p = 0.029, p = 0.0002), while expression of IGFBP-5, -rP2 (CTGF), -rP4 (Cyr61), FOSL1, LOXL2, OAZ1 and CDK4 was not different between groups. In line with the gene expression we found increased cell culture supernatant concentrations of IGFBP-3 (p = 0.006) in emphysema.
These data support the hypothesis that premature aging of lung fibroblasts occurs in emphysema, via a telomere-independent mechanism. The upregulation of the senescence-associated IGFBP-3 and -rP1 in emphysema suggests that inhibition of the action of insulin and insulin-like growth factors could be involved in the reduced
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Holz O, Zühlke I, Jaksztat E, Müller KC, Welker L, Nakashima M, Diemel KD, Branscheid D, Magnussen H, Jörres RA: Lung fibroblasts from patients with emphysema show a reduced proliferation rate in culture. Eur Respir J 2004, 24:575–579.
Nobukuni S, Watanabe K, Inoue J, Wen FQ, Tamaru N, Yoshida M: Cigarette smoke inhibits the growth of lung fibroblasts from patients with pulmonary emphysema. Respirology 2002, 7:217–223.
Noordhoek JA, Postma DS, Chong LL, Vos JT, Kauffman HF, Timens W, Van Straaten JF: Different proliferative capacity of lung fibroblasts obtained from control subjects and patients with emphysema. Exp Lung Res 2003, 29:291–302.
Bird J, Ostler EL, Faragher RG: Can we say that senescent cells cause ageing? Exp Gerontol 2003, 38:1319–1326.
Balin AK, Fisher AJ, Anzelone M, Leong I, Allen RG: Effects of establishing cell cultures and cell culture conditions on the proliferative life span of human fibroblasts isolated from different tissues and donors of different ages. Exp Cell Res 2002, 274:275–287.
Ogryzko VV, Hirai TH, Russanova VR, Barbie DA, Howard BH: Human fibroblast commitment to a senescence-like state in response to histone deacetylase inhibitors is cell cycle dependent. Mol Cell Biol 1996, 16:5210–5218.
Moodie FM, Marwick JA, Anderson CS, Szulakowski P, Biswas SK, Bauter MR, Kilty I, Rahman I: Oxidative stress and cigarette smoke alter chromatin remodeling but differentially regulate NF-kappaB activation and proinflammatory cytokine release in alveolar epithelial cells. FASEB J 2004, 18:1897–1899.
Absher M: Fibroblasts. In Lung Cell Biology. Lung Biology in Health and Disease. Volume 41. Edited by: Massaro D, Marcel Dekker. Inc New York, Basel; 1995:401–439.
Dimri GP, Lee X, Basile G, Acosta M, Scott G, Roskelley C, Medrano EE, Linskens M, Rubelj I, Pereira-Smith O: A biomarker that identifies senescent human cells in culture and in aging skin in vivo. Proc Natl Acad Sci U S A 1995, 92:9363–9367.
Kurz DJ, Decary S, Hong Y, Erusalimsky JD: Senescence-associated (beta)-galactosidase reflects an increase in lysosomal mass during replicative ageing of human endothelial cells. J Cell Sci 2000,113(Pt 20):3613–3622.
Severino J, Allen RG, Balin S, Balin A, Cristofalo VJ: Is beta-galactosidase staining a marker of senescence in vitro and in vivo? Exp Cell Res 2000, 257:162–171.
Harley CB, Futcher AB, Greider CW: Telomeres shorten during ageing of human fibroblasts. Nature 1990, 345:458–460.
Allsopp RC, Vaziri H, Patterson C, Goldstein S, Younglai EV, Futcher AB, Greider CW, Harley CB: Telomere length predicts replicative capacity of human fibroblasts. Proc Natl Acad Sci U S A 1992, 89:10114–10118.
Goldstein S, Moerman EJ, Jones RA, Baxter RC: Insulin-like growth factor binding protein 3 accumulates to high levels in culture medium of senescent and quiescent human fibroblasts. Proc Natl Acad Sci U S A 1991, 88:9680–9684.
Wilson HM, Birnbaum RS, Poot M, Quinn LS, Swisshelm K: Insulin-like growth factor binding protein-related protein 1 inhibits proliferation of MCF-7 breast cancer cells via a senescence-like mechanism. Cell Growth Differ 2002, 13:205–213.
Swisshelm K, Ryan K, Tsuchiya K, Sager R: Enhanced expression of an insulin growth factor-like binding protein (mac25) in senescent human mammary epithelial cells and induced expression with retinoic acid. Proc Natl Acad Sci U S A 1995, 92:4472–4476.
Pilewski JM, Liu L, Henry AC, Knauer AV, Feghali-Bostwick CA: Insulin-like growth factor binding proteins 3 and 5 are overexpressed in idiopathic pulmonary fibrosis and contribute to extracellular matrix deposition. Am J Pathol 2005, 166:399–407.
Schuller AG, van Neck JW, Beukenholdt RW, Zwarthoff EC, Drop SL: IGF, type I IGF receptor and IGF-binding protein mRNA expression in the developing mouse lung. J Mol Endocrinol 1995, 14:349–355.
Mohan S, Baylink DJ: Serum insulin-like growth factor binding protein (IGFBP)-4 and IGFBP-5 levels in aging and age-associated diseases. Endocrine 1997, 7:87–91.
Spira A, Beane J, Pinto-Plata V, Kadar A, Liu G, Shah V, Celli B, Brody JS: Gene expression profiling of human lung tissue from smokers with severe emphysema. Am J Respir Cell Mol Biol 2004, 31:601–610.
Ning W, Li CJ, Kaminski N, Feghali-Bostwick CA, Alber SM, Di YP, Otterbein SL, Song R, Hayashi S, Zhou Z, et al.: Comprehensive gene expression profiles reveal pathways related to the pathogenesis of chronic obstructive pulmonary disease. Proc Natl Acad Sci U S A 2004, 101:14895–14900.
Zhang Q, Adiseshaiah P, Reddy SP: Matrix metalloproteinase/epidermal growth factor receptor/mitogen-activated protein kinase signaling regulate fra-1 induction by cigarette smoke in lung epithelial cells. Am J Respir Cell Mol Biol 2005, 32:72–81.
Csiszar K: Lysyl oxidases: a novel multifunctional amine oxidase family. Prog Nucleic Acid Res Mol Biol 2001, 70:1–32.
Newman RM, Mobascher A, Mangold U, Koike C, Diah S, Schmidt M, Finley D, Zetter BR: Antizyme targets cyclin D1 for degradation. A novel mechanism for cell growth repression. J Biol Chem 2004, 279:41504–41511.
Erpenbeck VJ, Hohlfeld JM, Volkmann B, Hagenberg A, Geldmacher H, Braun A, Krug N: Segmental allergen challenge in patients with atopic asthma leads to increased IL-9 expression in bronchoalveolar lavage fluid lymphocytes. J Allergy Clin Immunol 2003, 111:1319–1327.
Smith PK, Krohn RI, Hermanson GT, Mallia AK, Gartner FH, Provenzano MD, Fujimoto EK, Goeke NM, Olson BJ, Klenk DC: Measurement of protein using bicinchoninic acid. Anal Biochem 1985, 150:76–85.
Kurozumi M, Matsushita T, Hosokawa M, Takeda T: Age-related changes in lung structure and function in the senescence-accelerated mouse (SAM): SAM-P/1 as a new murine model of senile hyperinflation of lung. Am J Respir Crit Care Med 1994, 149:776–782.
Valdes AM, Andrew T, Gardner JP, Kimura M, Oelsner E, Cherkas LF, Aviv A, Spector TD: Obesity, cigarette smoking, and telomere length in women. The Lancet 2005. DOI:10.1016/S0140–673(05)66630–5
Pascal T, Debacq-Chainiaux F, Chretien A, Bastin C, Dabee AF, Bertholet V, Remacle J, Toussaint O: Comparison of replicative senescence and stress-induced premature senescence combining differential display and low-density DNA arrays. FEBS Lett 2005, 579:3651–3659.
Aston C, Jagirdar J, Lee TC, Hur T, Hintz RL, Rom WN: Enhanced insulin-like growth factor molecules in idiopathic pulmonary fibrosis. Am J Respir Crit Care Med 1995, 151:1597–1603.
Hwa V, Oh Y, Rosenfeld RG: The insulin-like growth factor-binding protein (IGFBP) superfamily. Endocr Rev 1999, 20:761–787.
King GL, Kahn CR, Rechler MM, Nissley SP: Direct demonstration of separate receptors for growth and metabolic activities of insulin and multiplication-stimulating activity (an insulinlike growth factor) using antibodies to the insulin receptor. J Clin Invest 1980, 66:130–140.
Goldstein RH, Poliks CF, Pilch PF, Smith BD, Fine A: Stimulation of collagen formation by insulin and insulin-like growth factor I in cultures of human lung fibroblasts. Endocrinology 1989, 124:964–970.
Lucibello FC, Sewing A, Brusselbach S, Burger C, Muller R: Deregulation of cyclins D1 and E and suppression of cdk2 and cdk4 in senescent human fibroblasts. J Cell Sci 1993,105(Pt 1):123–133.
Quanjer PH, Tammeling GJ, Cotes JE, Pedersen OF, Peslin R, Yernault JC: Lung volumes and forced ventilatory flows. Report Working Party Standardization of Lung Function Tests, European Community for Steel and Coal. Official Statement of the European Respiratory Society. Eur Respir J Suppl 1993, 16:5–40.