The Lipoxygenase Inhibitor, Baicalein, Modulates Cell Adhesion and Migration by Up-regulation of Integrins and Vinculin in Rat Heart Endothelial Cells

Overview

Journal Br J Pharmacol

Publisher Wiley

Specialty Pharmacology

Date 2007 Jun 27

PMID 17592510

Citations 4

Authors

Y-C S Hsieh

S-J Hsieh

Y-S Chang

C-M Hsueh

S-L Hsu

Affiliations

Soon will be listed here.

Abstract

Background And Purpose: Endothelial cell proliferation, migration and adhesion are necessary for the formation of new blood vessels. We reported previously that baicalein strongly inhibited proliferation of rat heart endothelial cells and here we assess effects on migration and adhesion of these cells.

Experimental Approach: Effects of baicalein on endothelial migration and adhesion were determined by in vitro wound assays and in modified Boyden chambers. Protein expression and subcellular distribution in rat heart endothelial cells were analysed by immunoblots and immunofluorescence staining.

Results: Pretreatment with baicalein for 48 h resulted in a concentration-dependent inhibition of endothelial migration, with an IC(50) of approximately 20 microM. Adhesion assays revealed that baicalein stimulated endothelial cell adhesion to fibronectin and vitronectin, effects blocked by the synthetic peptide Arg-Gly-Asp (RGD). Moreover, treatment with a blocking antibody against integrin alpha5beta1 drastically attenuated baicalein-mediated endothelial adhesion to fibronectin, but not to vitronectin. Furthermore, baicalein-mediated anti-migration effect and adhesion promotion could be partially reversed by the addition of 12(S)-hydroxyeicosatetraenoic acid (12(S)-HETE). Western blot analysis indicated that baicalein increased expression levels of integrin-alpha5beta1, -alphavbeta3 and vinculin proteins. Immunofluorescence staining showed that baicalein induced a marked reorganization of actin stress fibres and the recruitment of vinculin and integrins to focal adhesion plaques, with consequently increased formation of focal adhesion contacts.

Conclusions And Implications: Baicalein markedly inhibited the migration and enhanced the adhesion of rat heart endothelial cells, possibly by up-regulation of the integrins (alpha5beta1 and alphavbeta3) and vinculin and by promotion of actin reorganization and focal adhesion contact formation.

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References

Mauro L, Sisci D, Bartucci M, Salerno M, Kim J, Tam T . SHC-alpha5beta1 integrin interactions regulate breast cancer cell adhesion and motility. Exp Cell Res. 1999; 252(2):439-48. DOI: 10.1006/excr.1999.4639. View

Hsu S, Hsieh Y, Hsieh W, Chou C . Baicalein induces a dual growth arrest by modulating multiple cell cycle regulatory molecules. Eur J Pharmacol. 2001; 425(3):165-71. DOI: 10.1016/s0014-2999(01)01144-x. View

Huang Y, Tsang S, Yao X, Chen Z . Biological properties of baicalein in cardiovascular system. Curr Drug Targets Cardiovasc Haematol Disord. 2005; 5(2):177-84. DOI: 10.2174/1568006043586206. View

Romer L, Birukov K, Garcia J . Focal adhesions: paradigm for a signaling nexus. Circ Res. 2006; 98(5):606-16. DOI: 10.1161/01.RES.0000207408.31270.db. View

Zeng Z, Jia Y, Hahn N, Markwart S, Rockwood K, Livant D . Role of focal adhesion kinase and phosphatidylinositol 3'-kinase in integrin fibronectin receptor-mediated, matrix metalloproteinase-1-dependent invasion by metastatic prostate cancer cells. Cancer Res. 2006; 66(16):8091-9. DOI: 10.1158/0008-5472.CAN-05-4400. View

Liu S . Radiolabeled multimeric cyclic RGD peptides as integrin alphavbeta3 targeted radiotracers for tumor imaging. Mol Pharm. 2006; 3(5):472-87. DOI: 10.1021/mp060049x. View

Saunders W, Bohnsack B, Faske J, Anthis N, Bayless K, Hirschi K . Coregulation of vascular tube stabilization by endothelial cell TIMP-2 and pericyte TIMP-3. J Cell Biol. 2006; 175(1):179-91. PMC: 2064509. DOI: 10.1083/jcb.200603176. View

Hynes R . Integrins: bidirectional, allosteric signaling machines. Cell. 2002; 110(6):673-87. DOI: 10.1016/s0092-8674(02)00971-6. View

Schuppan D, Popov Y, Porov Y . Hepatic fibrosis: from bench to bedside. J Gastroenterol Hepatol. 2002; 17 Suppl 3:S300-5. DOI: 10.1046/j.1440-1746.17.s3.18.x. View

10.

Sadik C, Sies H, Schewe T . Inhibition of 15-lipoxygenases by flavonoids: structure-activity relations and mode of action. Biochem Pharmacol. 2003; 65(5):773-81. DOI: 10.1016/s0006-2952(02)01621-0. View

11.

Liu J, Huang T, Cheng W, Lu F . Baicalein and baicalin are potent inhibitors of angiogenesis: Inhibition of endothelial cell proliferation, migration and differentiation. Int J Cancer. 2003; 106(4):559-565. DOI: 10.1002/ijc.11267. View

12.

Tsuruta D, Jones J . The vimentin cytoskeleton regulates focal contact size and adhesion of endothelial cells subjected to shear stress. J Cell Sci. 2003; 116(Pt 24):4977-84. DOI: 10.1242/jcs.00823. View

13.

Fu Y, Chi C, Yin S, Hwang B, Chiu Y, Hsu S . Norepinephrine induces apoptosis in neonatal rat endothelial cells via down-regulation of Bcl-2 and activation of beta-adrenergic and caspase-2 pathways. Cardiovasc Res. 2004; 61(1):143-51. DOI: 10.1016/j.cardiores.2003.10.014. View

14.

Sekiya K, Okuda H . Selective inhibition of platelet lipoxygenase by baicalein. Biochem Biophys Res Commun. 1982; 105(3):1090-5. DOI: 10.1016/0006-291x(82)91081-6. View

15.

Burridge K, Connell L . Talin: a cytoskeletal component concentrated in adhesion plaques and other sites of actin-membrane interaction. Cell Motil. 1983; 3(5-6):405-17. DOI: 10.1002/cm.970030509. View

16.

Pierschbacher M, Ruoslahti E . Cell attachment activity of fibronectin can be duplicated by small synthetic fragments of the molecule. Nature. 1984; 309(5963):30-3. DOI: 10.1038/309030a0. View

17.

Geiger B, Volk T, Volberg T . Molecular heterogeneity of adherens junctions. J Cell Biol. 1985; 101(4):1523-31. PMC: 2113916. DOI: 10.1083/jcb.101.4.1523. View

18.

Folkman J, Klagsbrun M . Angiogenic factors. Science. 1987; 235(4787):442-7. DOI: 10.1126/science.2432664. View

19.

Dejana E, Colella S, Languino L, Balconi G, Corbascio G, Marchisio P . Fibrinogen induces adhesion, spreading, and microfilament organization of human endothelial cells in vitro. J Cell Biol. 1987; 104(5):1403-11. PMC: 2114477. DOI: 10.1083/jcb.104.5.1403. View

20.

Cho H, Ueda M, Tamaoka M, Hamaguchi M, Aisaka K, Kiso Y . Novel caffeic acid derivatives: extremely potent inhibitors of 12-lipoxygenase. J Med Chem. 1991; 34(4):1503-5. DOI: 10.1021/jm00108a039. View