Ukr.Biochem.J. 2016; Volume 88, Issue 4, Jul-Aug, pp. 12-19


The mechanism of VEGF-mediated endothelial cells survival and proliferation in conditions of unfed-culture

T. V. Nikolaienko, V. V. Nikulina, D. V. Shelest, L. V. Garmanchuk

Educational and Scientific Centre “Institute of Biology”,
Taras Shevchenko National University of Kyiv, Ukraine;

The mechanisms of VEGF-mediated effects on endothelial cells during cancer development and progression is not clear. In present study the biological effects of VEGF, VEGF-rich culture medium of peritoneal macrophages from mice with Lewis lung carcinoma were studied on MAEC cell line under conditions of unfed culture. We have shown that VEGF increased cell proliferation by the 5th day of culturing vs control and anti-VEGF-treated cells. This effect was associated with increased consumption of glucose and NO production by the 2nd day while decreased – on the 5th day of cell culturing. VEGF-mediated NO production was dependent on Ca2+ ions. Block of Ca2+-channels (LaCl3) had more pronounced inhibitory effect vs chelator of Ca2+ ions (EDTA).  It was shown that peritoneal macrophages are the main suppliers of VEGF at tumor angiogenesis, as evidenced by the data obtained on model system of endothelial cells synchronized in G0/G1 phase.

Keywords: , , , , ,


  1. Arutyunyan I, Fatkhudinov T, Kananykhina E, Usman N, Elchaninov A, Makarov A, Bolshakova G, Goldshtein D, Sukhikh G. Role of VEGF-A in angiogenesis promoted by umbilical cord-derived mesenchymal stromal/stem cells: in vitro study. Stem Cell Res Ther. 2016 Mar 22;7:46. PubMed, PubMedCentral, CrossRef
  2. Szabo E, Schneider H, Seystahl K, Rushing EJ, Herting F, Weidner KM, Weller M. Autocrine VEGFR1 and VEGFR2 signaling promotes survival in human glioblastoma models in vitro and in vivo. Neuro Oncol. 2016 Mar 23. pii: now043. PubMed, CrossRef
  3. Yeh WL, Lin CJ, Fu WM. Enhancement of glucose transporter expression of brain endothelial cells by vascular endothelial growth factor derived from glioma exposed to hypoxia. Mol Pharmacol. 2008 Jan;73(1):170-7.  PubMed, CrossRef
  4. Mann GE, Yudilevich DL, Sobrevia L. Regulation of amino acid and glucose transporters in endothelial and smooth muscle cells. Physiol Rev. 2003 Jan;83(1):183-252. Review. PubMed, CrossRef
  5. Ezhilarasan R, Mohanam I, Govindarajan K, Mohanam S. Glioma cells suppress hypoxia-induced endothelial cell apoptosis and promote the angiogenic process. Int J Oncol. 2007 Mar;30(3):701-7. PubMed, PubMedCentral, CrossRef
  6. Babaei S, Stewart DJ. Overexpression of endothelial NO synthase induces angiogenesis in a co-culture model. Cardiovasc Res. 2002 Jul;55(1):190-200. PubMed, CrossRef
  7. Ono M. Molecular links between tumor angiogenesis and inflammation: inflammatory stimuli of macrophages and cancer cells as targets for therapeutic strategy. Cancer Sci. 2008 Aug;99(8):1501-6. Review. PubMed, CrossRef
  8. Khan MA, Assiri AM, Broering DC. Complement and macrophage crosstalk during process of angiogenesis in tumor progression. J Biomed Sci. 2015 Jul 22;22:58. Review.
    PubMed, PubMedCentral, CrossRef
  9. Rodrigues SF, Granger DN. Blood cells and endothelial barrier function. Tissue Barriers. 2015 Apr 3;3(1-2):e978720. PubMed, PubMedCentral, CrossRef
  10. Bastaki M, Nelli EE, Dell’Era P, Rusnati M, Molinari-Tosatti MP, Parolini S, Auerbach R, Ruco LP, Possati L, Presta M. Basic fibroblast growth factor-induced angiogenic phenotype in mouse endothelium. A study of aortic and microvascular endothelial cell lines. Arterioscler Thromb Vasc Biol. 1997 Mar;17(3):454-64. PubMed, CrossRef
  11. Pietrangeli CE, Skamene E, Edelson PJ, Kongshavn PA. Measurement of 5′-nucleotidase in mouse peritoneal macrophages in listeriosis. Infect Immun. 1981 Jun;32(3):1206-10. PubMed, PubMedCentral
  12. Garmanchouk LV, Pyaskovskaya ON, Solyanik GI. Influence of pro-angiogenic cytokines on proliferative activity and survival of endothelial cells. Biopolym Cell. 2010; 26(3):187-193. CrossRef
  13. Mosmann T. Rapid colorimetric assay for cellular growth and survival: application to proliferation and cytotoxicity assays. J Immunol Methods. 1983 Dec 16;65(1-2):55-63. PubMed, CrossRef
  14. Nicoletti I, Migliorati G, Pagliacci MC, Grignani F, Riccardi C. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J Immunol Methods. 1991 Jun 3;139(2):271-9. PubMed, CrossRef
  15. Nikolaienko Т, Petruk N, Shelest D, Garmanchuk L. Influence of VEGF, EGF and their antagonists on proliferative activity and glucose consumption by endothelial cells. Bull Shevchenko Nat Univ Kyiv. 2015; 1(69): 36-38.
  16. Green LC, Wagner DA, Glogowski J, Skipper PL, Wishnok JS, Tannenbaum SR. Analysis of nitrate, nitrite, and [15N]nitrate in biological fluids. Anal Biochem. 1982 Oct;126(1):131-8. PubMed, CrossRef
  17. Conn G, Soderman DD, Schaeffer MT, Wile M, Hatcher VB, Thomas KA. Purification of a glycoprotein vascular endothelial cell mitogen from a rat glioma-derived cell line. Proc Natl Acad Sci USA. 1990 Feb;87(4):1323-7. PubMed, PubMedCentral, CrossRef
  18. Carmeliet P, Jain RK. Principles and mechanisms of vessel normalization for cancer and other angiogenic diseases. Nat Rev Drug Discov. 2011 Jun;10(6):417-27. Review. PubMed, CrossRef
  19. Dvorak HF. Tumor Stroma, Tumor Blood Vessels, and Antiangiogenesis Therapy. Cancer J. 2015 Jul-Aug;21(4):237-43. Review. PubMed, CrossRef
  20. Polet F, Feron O. Endothelial cell metabolism and tumour angiogenesis: glucose and glutamine as essential fuels and lactate as the driving force. J Intern Med. 2013 Feb;273(2):156-65. Review. PubMed, CrossRef
  21. Kimura H, Ogura T, Kurashima Y, Weisz A, Esumi H. Effects of nitric oxide donors on vascular endothelial growth factor gene induction. Biochem Biophys Res Commun. 2002 Aug 30;296(4):976-82. PubMed, CrossRef
  22. Fleming I, Fisslthaler B, Dimmeler S, Kemp BE, Busse R. Phosphorylation of Thr(495) regulates Ca(2+)/calmodulin-dependent endothelial nitric oxide synthase activity. Circ Res. 2001 Jun 8;88(11):E68-75. PubMed, CrossRef

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