Abstract:【Objective】To investigate the expression of the extracellular matrix (ECM) on focal adhesion (FAs) protein paxillin and vinculin in cultured vascular smooth muscle cells (SMCs).【Methods】 Rat aortic SMCs of fourth~sixth generation were grown on coverslips coated with fibronectin ( FN) or laminin (LN),or planted on Matrigel for cultivation. The expression of paxillin and vinculin was detected by confocal immunofluorescence technique, and was demostrated by the double fluorescence staining of the FAs vinculin/F-actin and paxillin/F-actin.【Results】The vinculin expression planted on FN was the lowest, and the LN was the second, and the highest expression was in Matrigel ; the cytoplasm of the vinculin on FN was more distributed ,but less on FAs; and vinculin, which grew on the LN matrix and the Matrigel, were more localized in the FAs, but less distributed in the cytoplasm. The fluorescence intensity of Vinculin was (64.72±7.18)AU/μm2,(89.82±5.11)AU/μm2,(117.19±16.99)AU/μm2 respectively in FN, LN and Matrigel, which were in turn increased, and there was statistical significance between the groups (P<0.05).The paxillin grown on FN was the highest, while that on the LN came the second, and that on Matrigel showed the lowest expression; Paxillin in Matrigel was mainly distributed in FAs, and that on FN also got a higher expression in the cytoplasm, except for that distributed on FAs. The fluorescence intensity of Paxillin was(129.87±10.22)AU/μm2,(86.59±9.90)AU/μm2,(56.32±7.54)AU/μm2,respectively in FN, LN and Matrigel, which were in turn decreased, and there was statistical significance between the groups (P<0.05).【Conclusion】Different ECM components have effect on the expression of FAs molecule: FN can up regulate the expression of paxillin and down regulate the expression of vinculin; while the LN and Matrigel can increase the expression of vinculin and decrease the expression of paxillin.
何谢玲,蔡维君,刘敏,关莹露. 细胞外基质对血管平滑肌细胞桩蛋白和纽蛋白表达的影响[J]. 医学临床研究, 2016, 33(11): 2084-2087.
HE Xie-ling, CAI Wei-jun, LIU Min,et al. Effect of Extracellular Matrix Expression on Paxillin and Vinculin in Cultured Vascular Smooth Muscle Cells. JOURNAL OF CLINICAL RESEARCH, 2016, 33(11): 2084-2087.
[1] Cai W, Vosschulte R, Afsah-Hedjri A,et al. Altered balance between extracellular proteolysis and antiproteolysis is associated with adaptive coronary arteriogenesis[J].J Mol Cell Cardiol,2000,32(6):997-1011. [2] Gomez D, Owens GK. Smooth muscle cell phenotypic switching in atherosclerosis[J].Cardiovasc Res,2012, 95(2):156-164. [3] Veith C, Marsh LM, Wygrecka M, et al. Paxillin regulates pulmonary arterial smooth muscle cell function in pulmonary hypertension[J].Am J Pathol,2012, 181(5):1621-1633. [4] Ivaska J . Unanchoring integrins in focal adhesions[J].Nat Cell Biol,2012,14(10):981-983. [5] Hayward IP, Bridle KR, Campbell GR, et al. Effect of extracellular matrix proteins on vascular smooth muscle cell phenotype[J].Cell Biol Int,1995,19(10): 839-846. [6] Thyberg J, Hultgardh-Nilsson A. Fibronectin and the basement membrane components laminin and collagen type IV influence the phenotypic properties of subcultured rat aortic smooth muscle cells differently[J].Cell Tissue Res,1994,276(2): 263-271. [7] Chiang HY, Korshunov VA, Serour A, et al. Fibronectin is an important regulator of flow-induced vascular remodeling[J].Arterioscler Thromb Vasc Biol,2009, 29(7):1074-1079. [8] Worth NF, Rolfe BE, Song J, et al. Vascular smooth muscle cell phenotypic modulation in culture is associated with reorganisation of contractile and cytoskeletal proteins[J].Cell Motil Cytoskeleton,2001,49(3):130-145. [9] Subauste MC, Pertz O, Adamson ED, et al. Vinculin modulation of paxillin-FAK interactions regulates ERK to control survival and motility[J].J Cell Biol,2004, 165(3):371-381. [10] Cai W, Vosschulte R, Afsah-Hedjri A, et al. Altered balance between extracellular proteolysis and antiproteolysis is associated with adaptive coronary arteriogenesis[J].J Mol Cell Cardiol,2000,32(6):997-1011. [11] Paul A, Ben S, Devina J, et al. Mechanosenditive components of integrin adhesions: Role of vinculin[J].Exp Cell Res,2016,343(1):21-27. [12] Rajani K, Surinder KB, Frances EL, et al. FAK and paxillin, two potential targets in pancreatic cancer[J].Oncotarger,2016,7(21):31586-31601. [13] Cai WJ, Li MB, Wu X, et al. Activation of the integrins alpha 5beta 1 and alpha v beta 3 and focal adhesion kinase (FAK) during arteriogenesis[J].Mol Cell Biochem,2009, 322(1-2):161-169.