Ahmet Tarık BAYKAL, Betül BAYKAL, Müge SERHATLI, Zelal ADIGÜZEL, Mehmet Altuğ TUNÇER, Ömer KAÇAR

Proteomic evidence for the plasticity of cultured vascular smooth muscle cells

Morphological, functional, and gene expression studies have established the phenotypic plasticity of smooth muscle cells (SMCs). These cells have been shown to respond to environmental stimulants such as extracellular matrix and growth factors. Cellular changes can vary between extremes defined as the contractile and synthetic states. Various growth factors have been shown to have profound effects on the phenotype of these cells. In this study, we intended to investigate the effects of growth factor-rich medium on the protein expression of vascular SMCs in culture. Interestingly, transiently changing the type of medium did not result in any apparent morphological differences, yet we hypothesized that some cellular factors might still be altered. In order to understand what kind of intracellular molecular changes should be expected to occur during the medium change, we analyzed global protein expression changes using nano-LC-MS/MS in smooth muscle cell cultures that were isolated and grown in one medium formulation and temporarily switched to the other. Our data indicate that proteins playing a role in energy metabolism (glycolysis), translation, and folding of proteins are affected, along with regulatory molecules and cytoskeletal proteins. The individual proteins and their significance are discussed within the scope of this paper.

Anahtar Kelimeler:

Key words: Thoracic aortic aneurysm, label-free LC-MS, smooth muscle cell, pathway analysis, smooth muscle cell growth medium, DMEM/F12 growth medium

Proteomic evidence for the plasticity of cultured vascular smooth muscle cells

Keywords:

Key words: Thoracic aortic aneurysm, label-free LC-MS, smooth muscle cell, pathway analysis, smooth muscle cell growth medium, DMEM/F12 growth medium,

PDF

___

Eagle H. Nutrition needs of mammalian cells in tissue culture. Science 122: 501–14, 1955.
Amirkia V, Qiubao P. Cell-culture Database: Literature-based reference tool for human and mammalian experimentally based cell culture applications. Bioinformation 8: 237–8, 2012.
Owens GK, Kumar MS, Wamhoff BR. Molecular regulation of vascular smooth muscle cell differentiation in development and disease. Physiol Rev 84: 767–801, 2004.
Beamish JA, He P, Kottke-Marchant K et al. Molecular regulation of contractile smooth muscle cell phenotype: implications for vascular tissue engineering. Tissue Eng Pt B-Rev 16: 467–91, 2010.
Kumar MS, Owens GK. Combinatorial control of smooth muscle-specific gene expression. Arterioscl Throm Vas 23: 737–47, 2003.
Ailawadi G, Moehle CW, Pei H et al. Smooth muscle phenotypic modulation is an early event in aortic aneurysms. J Thorac Cardiovasc Surg 138: 1392–9, 2009.
Chamley-Campbell J, Campbell GR, Ross R. The smooth muscle cell in culture. Physiol Rev 59: 1–61, 1979.
Hayashi K, Saga H, Chimori Y et al. Differentiated phenotype of smooth muscle cells depends on signaling pathways through insulin-like growth factors and phosphatidylinositol 3-kinase. J Biol Chem 273: 28860–7, 1998.
Hedin U, Daum G, Clowes AW. Heparin inhibits thrombininduced mitogen-activated protein kinase signaling in arterial smooth muscle cells. J Vasc Surg 27: 512–20, 1998.
Pukac L, Huangpu J, Karnovsky MJ. Platelet-derived growth factor-BB, insulin-like growth factor-I, and phorbol ester activate different signaling pathways for stimulation of vascular smooth muscle cell migration. Exp Cell Res 242: 548–60, 1998.
Pukac LA, Carter JE, Ottlinger ME et al. Mechanisms of inhibition by heparin of PDGF stimulated MAP kinase activation in vascular smooth muscle cells. J Cell Physiol 172: 69–78, 1997.
Daum G, Hedin U, Wang Y et al. Diverse effects of heparin on mitogen-activated protein kinase-dependent signal transduction in vascular smooth muscle cells. Circ Res 81: 17–23, 1997.
Rauch BH, Millette E, Kenagy RD et al. Thrombin- and factor Xa-induced DNA synthesis is mediated by transactivation of fibroblast growth factor receptor-1 in human vascular smooth muscle cells. Circ Res 94: 340–5, 2004.
Louwrens HD, Kwaan HC, Pearce WH et al. Plasminogen activator and plasminogen activator inhibitor expression by normal and aneurysmal human aortic smooth muscle cells in culture. Eur J Vasc Endovasc 10: 289–93, 1995.
McMillan WD, Patterson BK, Keen RR et al. In situ localization and quantification of seventy-two-kilodalton type IV collagenase in aneurysmal, occlusive, and normal aorta. J Vasc Surg 22: 295– 305, 1995.
Cohen JR, Sarfati I, Danna D et al. Smooth muscle cell elastase, atherosclerosis, and abdominal aortic aneurysms. Ann Surg 216: 327–30, discussion 330–2, 1992.
Hayes IM, Jordan NJ, Towers S et al. Human vascular smooth muscle cells express receptors for CC chemokines. Arterioscl Throm Vas 18: 397–403, 1998.
Ray JL, Leach R, Herbert JM et al. Isolation of vascular smooth muscle cells from a single murine aorta. Methods Cell Sci 23: 185–8, 2001.
Dubey RK, Jackson EK, Gillespie DG et al. Catecholamines block the antimitogenic effect of estradiol on human coronary artery smooth muscle cells. J Clin Endocr Metab 89: 3922–31, 2004.
Mao D, Lee JK, VanVickle SJ et al. Expression of collagenase-3 (MMP-13) in human abdominal aortic aneurysms and vascular smooth muscle cells in culture. Biochem Bioph Res Co 261: 904– 10, 1999.
Phillippi JA, Klyachko EA, Kenny JP et al. Basal and oxidative stress-induced expression of metallothionein is decreased in ascending aortic aneurysms of bicuspid aortic valve patients. Circulation 119: 2498–506, 2009.
Walton LJ, Franklin IJ, Bayston T et al. Inhibition of prostaglandin E2 synthesis in abdominal aortic aneurysms: implications for smooth muscle cell viability, inflammatory processes, and the expansion of abdominal aortic aneurysms. Circulation 100: 48– 54, 1999.
Ito T, Ikeda U, Shimpo M et al. Serotonin increases interleukin-6 synthesis in human vascular smooth muscle cells. Circulation 102: 2522–7, 2000.
Acilan C, Serhatli M, Kacar O et al. Smooth muscle cells isolated from thoracic aortic aneurysms exhibit increased genomic damage, but similar tendency for apoptosis. DNA Cell Biol 31: 1523–34, 2012.
Leik CE, Willey A, Graham MF et al. Isolation and culture of arterial smooth muscle cells from human placenta. Hypertension 43: 837–40, 2004.
Akkoyunlu G, Üstünel İ, Demir R. Immunohistochemical distribution of transforming growth factor alpha, laminin, fibronectin and desmin in rat ovary. Turk J Biol 24: 467–81, 2000.
Hacariz O, Sayers G, Baykal AT. A proteomic approach to investigate the distribution and abundance of surface and internal Fasciola hepatica proteins during the chronic stage of natural liver fluke infection in cattle. J Proteome Res 11: 3592–604, 2012.
Cheng FY, Blackburn K, Lin YM et al. Absolute protein quantification by LC/MS(E) for global analysis of salicylic acid-induced plant protein secretion responses. J Proteome Res 8: 82–93, 2009.
Silva JC, Gorenstein MV, Li GZ et al. Absolute quantification of proteins by LCMSE: a virtue of parallel MS acquisition. Mol Cell Proteomics 5: 144–56, 2006.
Bostanci N, Heywood W, Mills K et al. Application of label-free absolute quantitative proteomics in human gingival crevicular fluid by LC/MS E (gingival exudatome). J Proteome Res 9: 2191–9, 2010.
D’Aguanno S, D’Alessandro A, Pieroni L et al. New insights into neuroblastoma cisplatin resistance: a comparative proteomic and meta-mining investigation. J Proteome Res 10: 416–28, 20 Vissers JP, Langridge JI, Aerts JM. Analysis and quantification of diagnostic serum markers and protein signatures for Gaucher disease. Mol Cell Proteomics 6: 755–66, 2007.
Pradel W, Mai R, Gedrange T et al. Cell passage and composition of culture medium effects proliferation and differentiation of human osteoblast-like cells from facial bone. J Physiol Pharmacol 59 (Suppl 5): 47–58, 2008.
Hill KL, Obrtlikova P, Alvarez DF et al. Human embryonic stem cell-derived vascular progenitor cells capable of endothelial and smooth muscle cell function. Exp Hematol 38: 246–57, 20 Kent L. Culture and maintenance of human embryonic stem cells. J Vis Exp 2009.
Kim HY, Kim MJ, Yang JE et al. Effects of intermittent media replacement on the gene expression of differentiating neural progenitor cells. Mol Biosyst 8: 602–8, 2012.
Ikeda K, Takahashi Y, Katagiri S. Effect of medium change on the development of in vitro matured and fertilized bovine oocytes cultured in medium containing amino acids. J Vet Med Sci 62: 121–3, 2000.
Boccardi C, Cecchettini A, Caselli A et al. A proteomic approach to the investigation of early events involved in vascular smooth muscle cell activation. Cell Tissue Res 328: 185–95, 2007.
Patton WF, Erdjument-Bromage H, Marks AR et al. Components of the protein synthesis and folding machinery are induced in vascular smooth muscle cells by hypertrophic and hyperplastic agents. Identification by comparative protein phenotyping and microsequencing. J Biol Chem 270: 21404– 10, 1995.
MacLellan DL, Steen H, Adam RM et al. A quantitative proteomic analysis of growth factor-induced compositional changes in lipid rafts of human smooth muscle cells. Proteomics 5: 4733–42, 2005.
Griffiths JB. The effect of medium changes on the growth and metabolism of the human diploid cell, W1-38. J Cell Sci 8: 43– 52, 1971.