Skip to main content

Advertisement

SpringerLink
Log in

Exploring the molecular mechanisms of OSU-03012 on vascular smooth muscle cell proliferation

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Restenosis is resulted from the proliferation and migration of vascular smooth muscle cells (VSMCs) from the arterial media into the intima within the vessel lumen following percutaneous transluminal coronary angioplasty (PTCA). OSU-03012, a synthetic compound (2-amino-N-{4-[5-(2-phenanthrenyl)-3-(trifluoromethyl)-1H-pyrazol-1-yl]-phenyl} acetamide) acting as a PDK-1 inhibitor, is used as an apoptosis-promoting anticancer drug. However, whether OSU-03012 can inhibit VSMC proliferation and migration following PTCA remains unclear. In this study, we used A10 smooth muscle cells cultured in 10% FBS for stimulating proliferation and evaluated the inhibitory effects of OSU-03012 on cell proliferation and migration. The data demonstrated that OSU-03012 dose-dependently inhibited A10 cell proliferation examined by Trypan blue, MTT and morphological alteration assays, and inhibited the levels of proliferation-related proteins, proliferating cell nuclear antigen (PCNA), phosphorylated ERK examined by western blotting. Additionally, 10 μM OSU-03012 also enhanced apoptosis examined using DAPI assay by regulating apoptosis-related proteins. Furthermore, compared with the control group, A10 cells treated with 10 μM OSU-03012 showed a lower number of migrating cells examined by Boyden Chamber assay, and a dose-dependently reduced NFkB-dependent and interferon-stimulated response element (ISRE) promoter luciferase activities, implying the anti-migration and anti-inflammation effects of OSU03012. Taken together, this study provides insights into the pharmacological mechanisms of OSU-03012 in preventing smooth muscle cell proliferation, migration, and inflammation supporting the novel discovery of OSU-03012 as an adjuvant therapy for balloon injury-induced restenosis.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6

Similar content being viewed by others

Abbreviations

VSMC:

Vascular smooth muscle cell

PTCA:

Ercutaneous transluminal coronary angioplasty

PCNA:

Proliferating cell nuclear antigen

ISRE:

Interferon-stimulated response element

References

  1. Isner JM, Pieczek A, Schainfeld R, Blair R, Haley L, Asahara T, Rosenfield K, Razvi S, Walsh K, Symes JF (1996) Clinical evidence of angiogenesis after arterial gene transfer of phVEGF165 in patient with ischaemic limb. Lancet 348:370–374

    Article  CAS  PubMed  Google Scholar 

  2. Serruys PW, Luijten HE, Beatt KJ, Geuskens R, de Feyter PJ, van den Brand M, Reiber JH, ten Katen HJ, van Es GA, Hugenholtz PG (1988) Incidence of restenosis after successful coronary angioplasty: a time-related phenomenon. A quantitative angiographic study in 342 consecutive patients at 1, 2, 3, and 4 months. Circulation 77:361–371

    CAS  PubMed  Google Scholar 

  3. Sturek M, Reddy HK (2002) New tools for prevention of restenosis could decrease the “oculo-stento” reflex. Cardiovasc Res 53:292–293

    Article  CAS  PubMed  Google Scholar 

  4. Zhu J, Huang JW, Tseng PH, Yang YT, Fowble J, Shiau CW, Shaw YJ, Kulp SK, Chen CS (2004) From the cyclooxygenase-2 inhibitor celecoxib to a novel class of 3-phosphoinositide-dependent protein kinase-1 inhibitors. Cancer Res 64:4309–4318

    Article  CAS  PubMed  Google Scholar 

  5. Yacoub A, Park MA, Hanna D, Hong Y, Mitchell C, Pandya AP, Harada H, Powis G, Chen CS, Koumenis C, Grant S, Dent P (2006) OSU-03012 promotes caspase-independent but PERK-, cathepsin B-, BID-, and AIF-dependent killing of transformed cells. Mol Pharmacol 70:589–603

    Article  CAS  PubMed  Google Scholar 

  6. Mosmann T (1983) Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods 65:55–63

    Article  CAS  PubMed  Google Scholar 

  7. Karim MA, Miller DD, Farrar MA, Eleftheriades E, Reddy BH, Breland CM, Samarel AM (1995) Histomorphometric and biochemical correlates of arterial procollagen gene expression during vascular repair after experimental angioplasty. Circulation 91:2049–2057

    CAS  PubMed  Google Scholar 

  8. Labinaz M, Pels K, Hoffert C, Aggarwal S, O’Brien ER (1999) Time course and importance of neoadventitial formation in arterial remodeling following balloon angioplasty of porcine coronary arteries. Cardiovasc Res 41:255–266

    Article  CAS  PubMed  Google Scholar 

  9. Jin G, Chieh-Hsi Wu J, Li YS, Hu YL, Shyy JY, Chien S (2000) Effects of active and negative mutants of Ras on rat arterial neointima formation. J Surg Res 94:124–132

    Article  CAS  PubMed  Google Scholar 

  10. Wu CH, Lin CS, Hung JS, Wu CJ, Lo PH, Jin G, Shyy YJ, Mao SJ, Chien S (2001) Inhibition of neointimal formation in porcine coronary artery by a Ras mutant. J Surg Res 99:100–106

    Article  CAS  PubMed  Google Scholar 

  11. Indolfi C, Chiariello M, Avvedimento EV (1996) Selective gene therapy for proliferative disorders: sense and antisense. Nat Med 2:634–635

    Article  CAS  PubMed  Google Scholar 

  12. Indolfi C, Coppola C, Torella D, Arcucci O, Chiariello M (1999) Gene therapy for restenosis after balloon angioplasty and stenting. Cardiol Rev 7:324–331

    Article  CAS  PubMed  Google Scholar 

  13. Fairman MP (1990) DNA polymerase delta/PCNA: actions and interactions. J Cell Sci 95(Pt 1):1–4

    CAS  PubMed  Google Scholar 

  14. Jonsson ZO, Hindges R, Hubscher U (1998) Regulation of DNA replication and repair proteins through interaction with the front side of proliferating cell nuclear antigen. EMBO J 17:2412–2425

    Article  CAS  PubMed  Google Scholar 

  15. Gomez Roig E, Vazquez-Ramos JM (2003) Maize DNA polymerase alpha is phosphorylated by a PCNA-associated cyclin/Cdk complex: effect of benzyladenine. J Plant Physiol 160:983–990

    Article  PubMed  Google Scholar 

  16. Wei GL, Krasinski K, Kearney M, Isner JM, Walsh K, Andres V (1997) Temporally and spatially coordinated expression of cell cycle regulatory factors after angioplasty. Circ Res 80:418–426

    CAS  PubMed  Google Scholar 

  17. Sakoguchi-Okada N, Takahashi-Yanaga F, Fukada K, Shiraishi F, Taba Y, Miwa Y, Morimoto S, Iida M, Sasaguri T (2007) Celecoxib inhibits the expression of survivin via the suppression of promoter activity in human colon cancer cells. Biochem Pharmacol 73:1318–1329

    Article  CAS  PubMed  Google Scholar 

  18. Zhang S, Suvannasankha A, Crean CD, White VL, Johnson A, Chen CS, Farag SS (2007) OSU-03012, a novel celecoxib derivative, is cytotoxic to myeloma cells and acts through multiple mechanisms. Clin Cancer Res 13:4750–4758

    Article  CAS  PubMed  Google Scholar 

  19. Bellas RE, Lee JS, Sonenshein GE (1995) Expression of a constitutive NF-kappa B-like activity is essential for proliferation of cultured bovine vascular smooth muscle cells. J Clin Invest 96:2521–2527

    Article  CAS  PubMed  Google Scholar 

  20. Selzman CH, Shames BD, Reznikov LL, Miller SA, Meng X, Barton HA, Werman A, Harken AH, Dinarello CA, Banerjee A (1999) Liposomal delivery of purified inhibitory-kappaBalpha inhibits tumor necrosis factor-alpha-induced human vascular smooth muscle proliferation. Circ Res 84:867–875

    CAS  PubMed  Google Scholar 

  21. Landry DB, Couper LL, Bryant SR, Lindner V (1997) Activation of the NF-kappa B and I kappa B system in smooth muscle cells after rat arterial injury. Induction of vascular cell adhesion molecule-1 and monocyte chemoattractant protein-1. Am J Pathol 151:1085–1095

    CAS  PubMed  Google Scholar 

  22. Wainwright CL, Miller AM, Wadsworth RM (2001) Inflammation as a key event in the development of neointima following vascular balloon injury. Clin Exp Pharmacol Physiol 28:891–895

    Article  CAS  PubMed  Google Scholar 

  23. Reid LE, Brasnett AH, Gilbert CS, Porter AC, Gewert DR, Stark GR, Kerr IM (1989) A single DNA response element can confer inducibility by both alpha- and gamma-interferons. Proc Natl Acad Sci USA 86:840–844

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgement

This study was supported by a grant from China Medical University (CMU94 -032, CMU95-028, CMU95-030, and CMU95-029) and from Taiwan Department of Health Clinical Trial and Research Center of Excellence (DOH99-TD-B-111-004) and in part by Taiwan Department of Health Cancer Research Center of Excellence (DOH99-TDC-111-005).

Conflict of interest statement

The authors have declared no conflict of interest statement.

Author information

Authors and Affiliations

  1. Department of Biological Science and Technology, College of Life Sciences, China Medical University, Taichung, Taiwan, ROC

    Wei-Wen Kuo, Jing-Ru Weng, Wei-Hung Liu & Shih-Chang Tsai

  2. Research Center for Biodiversity, China Medical University, Taichung, Taiwan, ROC

    Jing-Ru Weng

  3. Graduate Institute of Basic Medical Science, China Medical University, Taichung, Taiwan, ROC

    Chih-Yang Huang

  4. Department of Health and Nutrition Biotechnology, Asia University, Taichung, Taiwan, ROC

    Chih-Yang Huang

  5. Department of Healthcare Administration, Asia University, Taichung, Taiwan, ROC

    Chang-Hai Tsai

  6. Department of Biological Science and Technology, National Chiao Tung University, Hsinchu, Taiwan, ROC

    Cheng-Hao Wen

  7. Department of Pharmacology, School of Pharmacology, China Medical University, No. 91, Hsueh-Shih Road, Taichung, 40402, Taiwan, ROC

    Chieh-Hsi Wu

Authors
  1. Wei-Wen Kuo
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Jing-Ru Weng
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Chih-Yang Huang
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Chang-Hai Tsai
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. Wei-Hung Liu
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. Cheng-Hao Wen
    View author publications

    You can also search for this author in PubMed Google Scholar

  7. Shih-Chang Tsai
    View author publications

    You can also search for this author in PubMed Google Scholar

  8. Chieh-Hsi Wu
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Chieh-Hsi Wu.

Additional information

Wei-Wen Kuo, Jing-Ru Weng, and Chih-Yang Huang have contributed equally to this work.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Kuo, WW., Weng, JR., Huang, CY. et al. Exploring the molecular mechanisms of OSU-03012 on vascular smooth muscle cell proliferation. Mol Cell Biochem 344, 81–89 (2010). https://doi.org/10.1007/s11010-010-0531-5

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11010-010-0531-5

Keywords

Search

Navigation