The signal transduction pathways of heat shock protein 27 phosphorylation in vascular smooth muscle cells



The objective of this study is to investigate the signal transduction pathways that regulate heat shock protein 27 (HSP27) phosphorylation and migration of vascular smooth muscle cells (VSMCs) from spontaneously hypertensive rats (SHR) induced by angiotensin II (AngII) and platelet derived growth factor-BB (PDGF-BB). The activity of HSP27 was evaluated by Western blot with specific phospho-HSP27 antibody. F-actin polymerization was detected by FITC-Phalloidine staining using confocal microscopy. Modified Boyden chamber technique was employed for VSMCs migration assessment. Within a given concentration, the phosphorylation of HSP27 induced by AngII and PDGF-BB was blocked by the specific P38MAPK inhibitor SB202190, the specific PI3K inhibitor LY294002 and the specific ERK1/2 inhibitor U0126 in a concentration-dependent manner, with a peak inhibition rate at 87.2%, 78.4% and 37.3%, respectively, induced by AngII (P < 0.01), with a peak inhibition rate at 85.0%, 55.3% and 41.0%, respectively, induced by PDGF-BB (P < 0.01).The migration of VSMCs induced by AngII and PDGF-BB was inhibited by 100 μmol/l SB202190, 30 μmol/l LY294002, and 30 μmol/l U0126, with a inhibition rate at 60.1%, 71.7% and 47.3%, respectively, provoked by AngII (P < 0.01), with a inhibition rate at 55.3%, 55.6% and 38.1%, respectively, provoked by PDGF-BB (P < 0.01). P38MAPK and PI3 K/Akt are important pathways that contribute to the phosphorylation of HSP27 and migration of VSMCs in response to AngII and PDGF-BB. ERK1/2 might be involved in HSP27 phosphorylation and migration of VSMCs provoked by AngII and PDGF-BB.


Heat shock protein 27 Cytoskeleton Cell migration Vascular smooth muscle cells 


  1. 1.
    Chen HF, Xie LD, Xu CS (2009) Role of heat shock protein 27 phosphorylation in migration of vascular smooth muscle cells. Mol Cell Biochem 327(1-2):1–6CrossRefPubMedGoogle Scholar
  2. 2.
    Tartakover-Matalon S, Cherepnin N, Kuchuk M et al (2007) Impaired migration of trophoblast cells caused by simvastatin is associated with decreased membrane IGF-I receptor, MMP-2 activity and HSP27 expression. Hum Reprod 22(4):1161–1167CrossRefPubMedGoogle Scholar
  3. 3.
    Nadin SB, Vargas-Roig LM, Drago G et al (2007) Hsp27, Hsp70 and mismatch repair proteins Hmlh1 and Hmsh2 expression in peripheral blood lymphocytes from healthy subjects and cancer patients. Cancer Lett 252(1):131–146CrossRefPubMedGoogle Scholar
  4. 4.
    Jog NR, Jala VR, Ward RA et al (2007) Heat shock protein 27 regulates neutrophil chemotaxis and exocytosis through two independent mechanisms. J Immunol 178(4):2421–2428PubMedGoogle Scholar
  5. 5.
    Somara S, Bitar KN (2004) Tropomyosin interacts with phosphorylated HSP27 in agonist-induced contraction of smooth muscle. Am J Physiol Cell Physiol 286:C1290–C1301CrossRefPubMedGoogle Scholar
  6. 6.
    An SS, Fabry B, Mellema M et al (2003) Role of heat shock protein 27 in cytoskeletal remodeling of the airway smooth muscle cell. J Appl Physiol 96:1701–1713CrossRefGoogle Scholar
  7. 7.
    Cao H, Dronadula N, Rizvi F et al (2006) Novel role for STAT-5B in the regulation of Hsp27-FGF-2 axis facilitating thrombin-induced vascular smooth muscle cell growth and motility. Circ Res 98:913–922CrossRefPubMedGoogle Scholar
  8. 8.
    Meier M, King GL, Clermont A et al (2001) Angiotensin AT1 receptor stimulates heat shock protein 27 phosphorylation in vitro and in vivo. Hypertension 38:1260–1265CrossRefPubMedGoogle Scholar
  9. 9.
    Chen Y, Ross BM, Currie RW (2004) Heat shock treatment protects against angiotensin II-induced hypertension and inflammation in aorta. Cell Stress Chaperones 9:99–107PubMedGoogle Scholar
  10. 10.
    Chen Y, Currie RW (2006) Small interfering RNA knocks down heat shock factor-1(HSF-1) and exacerbates pro-inflammatory activation of NF-kB and AP-1 in vascular smooth muscle cells. Cardiovascular Res 69:66–75CrossRefGoogle Scholar
  11. 11.
    O’Shaughnessy RF, Welti JC, Cooke JC et al (2007) AKT-dependent HspB1 (Hsp27) activity in epidermal differentiation. J Biol Chem 282(23):17297–17305CrossRefPubMedGoogle Scholar
  12. 12.
    Peart JN, Gross ER, Headrick JP et al (2007) Impaired p38 MAPK/HSP27 signaling underlies aging-related failure in opioid-mediated cardioprotection. J Mol Cell Cardiol 42(5):972–980CrossRefPubMedGoogle Scholar
  13. 13.
    Zheng C, Lin Z, Zhao ZJ et al (2006) MAPK-activated protein kinase-2 (MK2)-mediated formation and phosphorylation-regulated dissociation of the signal complex consisting of p38, MK2, Akt, and Hsp27. J Biol Chem 281(48):37215–37226CrossRefPubMedGoogle Scholar
  14. 14.
    Takai S, Tokuda H, Matsushima-Nishiwaki R et al (2006) Phosphatidylinositol 3-kinase/Akt plays a role in sphingosine 1-phosphate-stimulated HSP27 induction in osteoblasts. J Cell Biochem 98(5):1249–1256CrossRefPubMedGoogle Scholar
  15. 15.
    Rane MJ, Pan Y, Singh S et al (2003) Heat shock protein 27 controls apoptosis by regulating Akt activation. J Biol Chem 278(30):27828–27835CrossRefPubMedGoogle Scholar
  16. 16.
    Hirade K, Tanabe K, Niwa M et al (2005) Adenylyl cyclase-cAMP system inhibits thrombin-induced HSP27 in vascular smooth muscle cells. J Cell Biochem 94:573–584CrossRefPubMedGoogle Scholar
  17. 17.
    Takenaka M, Matsuno H, Ishisaki A et al (2004) Platelet-derived growth factor-BB phosphorylates heat shock protein 27 in cardiac myocytes. J Cell Biochem 91:316–324CrossRefPubMedGoogle Scholar
  18. 18.
    Akamatsu S, Nakajima K, Ishisaki A et al (2004) Vasopressin phosphorylates HSP27 in aortic smooth muscle cells. J Cell Biochem 92:1203–1211CrossRefPubMedGoogle Scholar
  19. 19.
    Lee CK, Lee HM, Kim HJ et al (2007) Syk contributes to PDGF-BB-mediated migration of rat aortic smooth muscle cells via MAPK pathways. Cardiovasc Res 74(1):159–168CrossRefPubMedGoogle Scholar
  20. 20.
    Guo K, Liu Y, Zhou H et al (2008) Involvement of protein kinase C beta-extracellular signal-regulating kinase 1/2/p38 mitogen-activated protein kinase-heat shock protein 27 activation in hepatocellular carcinoma cell motility and invasion. Cancer Sci 99(3):486–496CrossRefPubMedGoogle Scholar
  21. 21.
    Madan P, Calder MD, Watson AJ et al (2005) Mitogen-activated protein kinase (MAPK) blockade of bovine preimplantation embryogenesis requires inhibition of both p38 and extracellular signal-regulated kinase (ERK) pathways. Reproduction 130(1):41–51CrossRefPubMedGoogle Scholar
  22. 22.
    Borbiev T, Verin AD, Birukova A et al (2003) Role of CaM kinase II and ERK activation in thrombin-induced endothelial cell barrier dysfunction. Am J Physiol Lung Cell Mol Physiol 285(1):L43–L54PubMedGoogle Scholar
  23. 23.
    Borbiev T, Verin AD, Shi S et al (2001) Regulation of endothelial cell barrier function by calcium/calmodulin-dependent protein kinase II. Am J Physiol Lung Cell Mol Physiol 280(5):L983–L990PubMedGoogle Scholar
  24. 24.
    Nguyen A, Chen P, Cai H (2004) Role of CaMKII in hydrogen peroxide activation of ERK1/2, p38 MAPK, HSP27 and actin reorganization in endothelial cells. FEBS Lett 572:307–313CrossRefPubMedGoogle Scholar
  25. 25.
    Pichon S, Bryckaert M, Berrou E (2004) Control of actin dynamics by p38 MAP kinase-HSP27 distribution in the lamellipodium of smooth muscle cells. J Cell Sci 117:2569–2577CrossRefPubMedGoogle Scholar
  26. 26.
    Meloche S, Landry J, Huot J et al (2000) P38 MAP kinase pathway regulates angiotensin II-induced contraction of rat vascular smooth muscle. Am J Physiol Heart Circ Physiol 279:H741–H751PubMedGoogle Scholar
  27. 27.
    Tanabe K, Akamatsu S, Suga H et al (2005) Midazolam suppresses thrombin-induced heat shock protein 27 phosphorylation through inhibition of p38 mitogen-activated protein kinase in cardiac myocytes. J Cell Biochem 96:56–64CrossRefPubMedGoogle Scholar
  28. 28.
    Hirade K, Kozawa O, Tanabe K et al (2002) Thrombin stimulates dissociation and induction of HSP27 via p38 MAPK in vascular smooth muscle cells. Am J Physiol Heart Circ Physiol 283:H941–H948PubMedGoogle Scholar
  29. 29.
    Nakajima K, Hirade K, Ishisaki A et al (2005) Akt regulates thrombin-induced HSP27 phosphorylation in aortic smooth muscle cells: function at a point downstream from p38 MAP Kinase. Life Sci 77:96–107CrossRefPubMedGoogle Scholar
  30. 30.
    Mearow KM, Dodge ME, Rahimtula M et al (2002) Stress-mediated signaling in PC12 cells—the role of the small heat shock protein, Hsp27, and Akt in protecting cells from heat stress and nerve growth factor withdrawal. J Neurochem 83(2):452–462CrossRefPubMedGoogle Scholar
  31. 31.
    Fukagawa Y, Nishikawa J, Kuramitsu Y et al (2008) Epstein-Barr virus upregulates phosphorylated heat shock protein 27 kDa in carcinoma cells using the phosphoinositide 3-kinase/Akt pathway. Electrophoresis 29(15):3192–3200PubMedGoogle Scholar
  32. 32.
    Havasi A, Li Z, Wang Z et al (2008) Hsp27 inhibits Bax activation and apoptosis via a phosphatidylinositol 3-kinase-dependent mechanism. J Biol Chem 283(18):12305–12313CrossRefPubMedGoogle Scholar
  33. 33.
    Rafiee P, Theriot ME, Nelson VM et al (2006) Human esophageal microvascular endothelial cells respond to acidic pH stress by PI3K/AKT and p38 MAPK-regulated induction of Hsp70 and Hsp27. Am J Physiol Cell Physiol 291(5):C931–C945CrossRefPubMedGoogle Scholar
  34. 34.
    Suga H, Nakajima K, Shu E et al (2005) Possible involvement of phosphatidylinositol 3-kinase/Akt signal pathway in vasopressin-induced HSP27 phosphorylation in aortic smooth muscle A10 cells. Arch Biochem Biophys 438:137–145CrossRefPubMedGoogle Scholar

Copyright information

© Springer Science+Business Media, LLC. 2009

Authors and Affiliations

  1. 1.Fujian Hypertension Research Institute, First Affiliated HospitalFujian Medical UniversityFuzhouChina
  2. 2.Department of Cardiology, Fujian Provincial Clinical CollegeFujian Medical UniversityFuzhouChina

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