Effect of ZNimesulide on the Differentiation and Survival of Endothelial Progenitor Cells

  • Oh, Ho-Kyun (Cancer Research Institute, Catholic Research of Medical Science, The Catholic University of Korea) ;
  • Kim, Sun-Yong (Cancer Research Institute, Catholic Research of Medical Science, The Catholic University of Korea) ;
  • Baek, Sang-Hong (Department of Internal Medicine, Division of Cardiovascular Medicine, The Catholic University of Korea) ;
  • Lim, Sung-Cil (Department of Internal Medicine, Division of Cardiovascular Medicine, The Catholic University of Korea) ;
  • Ahn, Hyun-Young (Department of Obstetrics and Gynecology, Division of Maternal-Fetal Medicine, Korea Institute of Radiological and Medical Sciences) ;
  • Shin, Jong-Chul (Department of Obstetrics and Gynecology, Division of Maternal-Fetal Medicine, Korea Institute of Radiological and Medical Sciences) ;
  • Hong, Sung-Hee (Laboratory of Experimental Therapeutics, Korea Institute of Radiological and Medical Sciences) ;
  • Hong, Yong-Kil (Cancer Research Institute, Catholic Research of Medical Science, The Catholic University of Korea) ;
  • Joe, Young-Ae (Cancer Research Institute, Catholic Research of Medical Science, The Catholic University of Korea)
  • Published : 2004.12.01


Nonsteroidal anti-inflammatory drugs (NSAIDs), particularly the highly selective cyclooxygenase (COX)-2 inhibitors have been shown to decrease the growth of tumor, in part, by inhibition of neovascularization. Recently, besides mature endothelial cells, endothelial progenitor cells (EPCs) have been shown to contribute neovascularization in angiogenic tissues. In this study, we addressed a question whether nimesulide, a selective COX-2 inhibitor, could affect differentiation of EPCs into adhesive endothelial cells in vitro. Total mononuclear cells were isolated from cord blood by Ficoll density gradient centrifugation, and then the cells were incubated with nimesulide or vehicle control for 7 days. The number of adherent and spindle-shaped cells decreased by nimesulide treatment in a concentration-dependent fashion at a concentration range of 5 - 200 ${\mu}M$. Moreover, the adherent cells double positive for DiI-ac-LDL uptake and lectin binding significantly decreased upon nimesulide treatment. There was no change of expression of CD31 between treatment and control groups, whereas slight reduction was detected upon treatment in expression of VE-cadherin, ICAM-1, vWF, ${\alpha}v$, and ${\alpha}5$. Nimesulide also reduced cell viability during first 3 days' culture and induced apoptosis in adherent EPCs, resulting in increased annexin-V-positive and propidium iodide-negative cells. Taken together, these results suggest that nimesulide could be applied for the inhibition of new vessel formation, in part, by inhibiting differentiation and survival of EPCs.



  1. Asahara, T., Murohara, T., Sullivan, M., van der Zee R., Li, T., Witzenbichler, B., Schatteman, G. and Isner, J. M. (1997). Isolation of putative progenitor endothelial cells for angiogenesis. Science. 275, 964-967 https://doi.org/10.1126/science.275.5302.964
  2. Asahara, T., Masuda, H., Takahashi, T., Kalka, c., Pastore, C., Siver, M., Kearne, M., Magner, M. and Isner, J. M. (1999). Bone marrow origin of endothelial progenitor cells responsible for postnatal vasculogenesis in physiological and pathological neovascularization. Cir. Res. 85, 221-228 https://doi.org/10.1161/01.RES.85.3.221
  3. Boyer, M., Townsend, L. E., Vogel, L.M., Falk, J., Reitz-Vick, D., Trevor, K. T., Villaba, M., Bendick, P.J. and Glover, J. L. (2000). Isolation of endothelial cells and their progenitor cells form human peripheral blood. J. Vase. Surg. 31, 181-189 https://doi.org/10.1016/S0741-5214(00)70080-2
  4. Bussolino, F., Mantovani, A. and Persico, G. (1997). Molecular mechanisms of blood vessel formation. Trends Biochem. Sci, 22,251-256 https://doi.org/10.1016/S0968-0004(97)01074-8
  5. Dominguez-Jimenez, C., Diaz-Gonalez, E, Gonzalez-Alvaro, I., Cesar, J. M. and Sanchez-Madrid, F. (1999). Prevention of $\alpha$II(b)$\beta$$_3$ activation by non-steroidal anti-inflammatory drugs. FEBS Lett. 446, 318-322 https://doi.org/10.1016/S0014-5793(99)00236-7
  6. Dormand, O., Foletti, A., Paroz, C. and Ruegg, C. (2001). NSAIDs inhibit $\alpha$v$\beta$$_3$ integrin-mediated and Cdc42/Racdependent endothelial-cell spreading, migration and angiogenesis. Nat. Med. 7,1041-1047 https://doi.org/10.1038/nm0901-1041
  7. Eibl, G., Bremmer, D., Okada, Y., Duffy, J. P., Law, R. E., Reber, H. A. and Hines, O. J. (2003). PGE$_2$ is generated by specific COX-2 activity and increases VEGF production in COX-2expressing human pancreatic cancer cells. Biochem. Biophys. Res. Commun. 306, 887-897 https://doi.org/10.1016/S0006-291X(03)01079-9
  8. Famaey, J. P. (1997). In vitro and in vivo pharmacological evidence of selective cyclooxygenas-2 inhibition by nimesulide: An overview. Inflamm. Res. 46, 437-446 https://doi.org/10.1007/s000110050221
  9. Fukutake, M., Nakatsugi, S., Isoi, T., Takahashi, M., Ohta, T., Mamiya, S., Taniguchi, Y., Sato, H., Fukuda, K., Sugimura, T. and Wakabayashi, K. (1998). Suppressive effects of nimesulide, a selective inhibitor of cyclooxygenase-2, on azoxymethaneinduced colon carcinogenesis in mice. Carcinogenesis. 19, 1939-1942 https://doi.org/10.1093/carcin/19.11.1939
  10. Garcia-Vicuna, R., Garcia-Vicuna, R., Diaz-Gonzalez, F., Gonzalez-Alvaro, I., del Pozo, M. A., Mollinedo, E, Cabanas, C., Gonzalez-Amaro, R. and Sanchez-Madrid, F. (1997). Prevention of cytokine-induced changes in leukocyte adheresion receptors by nonsteroidal anti-inflammatory drugs from the oxicam family. Arthritis Rheum. 40, 143-153 https://doi.org/10.1002/art.1780400119
  11. Giardiello, F. M., Hamilton, S. R., Krush, A. J., Piantadosi, S., Hylind, L. M., Celano, P., Booker, S. V., Robinson, C. R. and Offerhaus, G. J. (1993). Treatment of colonic and rectal adenomas with sulindac in familial adenomatous polyposis. N. Engl. J. Med. 328,1313-1316 https://doi.org/10.1056/NEJM199305063281805
  12. Hida, T., Kozaki, K. I., Muramatsu, H., Masuda, A., Shimizu, S.,Mitsudomi, T., Sugiura, T., Ogawa, M. and Takahashi, T. (2000). Cyclooxygenase-2 inhibitor induces apoptosis and enhances cytotoxity of various anticancer agents in non-small cell lung cancer cell lines. Clin. Cancer Res. 6, 2006-2011
  13. Joe, Y. A, Back, S. H., Park, H. Y., Lee, Y. H., Kwon, H. K., Kim, Y. J., Lee, S. Y., Seung, K. B., Chae, J. S., Kim, J. H., Hong, S. J. and Choi, K. B. (2002). In vitro differentiation of endothelial precursor cells derived from umbilical cord blood. Korea circulation J. 32, 646-654 https://doi.org/10.4070/kcj.2002.32.8.646
  14. Li, X. H., Li, J. J., Zhang, H. W., Sun, P., Zhang, Y. L., Cai, S. H. and Ren, X. D. (2003). Nimesulide inhibits tumor growth in mice implanted hepatoma: overexpression of Bax over Bcl-2. Acta Pharmacol Sin. 24, 1045-1050
  15. Murohara, T., Ikeda, H., Duan, J., Shintani, S., Sasaki, K., Eguchi, H., Onitsuka, I., Matsui; K. and Imaizumi, T. (2000). Transplanted cord blood-derived endothelial precursor cells augment postnatal neovasularization. J. Clin. Invest. 105, 1527-1536 https://doi.org/10.1172/JCI8296
  16. Risau, W. (1997). Mechanism of angiogenesis. Nature. 386, 671-674 https://doi.org/10.1038/386671a0
  17. Shaik, M. S., Chatterjee, A and Singh, M. (2004). Effect of a selective cyclooxygenase-2 inhibitor, nimesulide, on the growth of lung tumors and their expresstion of cyclooxygenae-2 and peroxisome proliferators-activated receptor-$\gamma$ Clin. Cancer Res.10, 1521-1529 https://doi.org/10.1158/1078-0432.CCR-0902-03
  18. Shi, Q., Rafii, S., Wu, M. H., Wijelath, E. S., Yu, C., Ishida, A., Fujit, Y., Kothari, S., Mohle, R., Sauvage, L. R., Moor, M. A., Storb, R. F. and Hammond, W. P. (1998). Evidence for circulating bone marrow-derived endothelial cells. Blood. 92, 362-367
  19. Shintani, S., Murohara, T., Ikeda H., Ueno, T., Honma, T., Katoh, A., Sasaki, K., Shimada, T., Oike, Y. and Imaizumi, T. (2001). Mobilization of endothelila progenitor cells in patients with acute myocardial infarction. Circulation. 103, 2776-2779 https://doi.org/10.1161/hc2301.092122
  20. Tsujii, M., Kawano, S., Tsuji, S., Sawaoka, H., Hori, M. and DuBois, R. N. (1998). cyclooxygenase regulates angiogenesis induced by colon cancer cells. Cell. 93,705-716 https://doi.org/10.1016/S0092-8674(00)81433-6
  21. Walter, D. H., Rittig, K., Bahlmann, F. H., Kirchmair, R., Silver, M., Murayama, T., Nishimura, H., Losordo, D. W., Asahara, T. and Isner, J. M. (2002). Statin Therapy Accelerates Reendothelialization A novel effect involving mobilization and incorporation of bone marrow-derived endothelilal progenitor cells. Circulation. 105, 3017-3024 https://doi.org/10.1161/01.CIR.0000018166.84319.55
  22. Warrington, S. J., Ravic, M. and Dawnay, A. (1993). Renal and general tolerability of repeated doses of nimesulide in normal subjects. Drugs. 46, 263-269
  23. Williams, C. S., Mann, M. and DuBois, R. N. (1999). The role of cyclooxygenases in inflammation, cancer, and development. Oncogene. 18,7908-7916 https://doi.org/10.1038/sj.onc.1203286
  24. Williams, C. S., Tsujii, M., Reese, J., Dey, S. K. and DuBois, R. N. (2000). Host cyclooxygenase-2 modulates carcinoma growth. J. Clin. Invest. 105, 1589-1594 https://doi.org/10.1172/JCI9621