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Dose-dependent effect of nocodazole on endothelial cell cytoskeleton

  • K. M. Smurova
  • A. A. Birukova
  • A. D. Verin
  • I. B. Alieva
Article
  • 573 Downloads

Abstract

The endothelium lining the inner surface of blood vessels fulfils an important barrier function and specifically, it controls vascular membrane permeability as well as nutrient and metabolite exchange in circulating blood and tissue fluids. Disturbances in vascular endothelium barrier function (vascular endothelium dysfunction) are coupled to cytoskeleton rearrangements, actomyosin contractility, and as a consequence, formation of paracellular gaps between endothelial cells. Microtubules constitute the first effector link in the reaction cascade resulting in vascular endothelium dysfunction. Increased vascular permeability associated with many human diseases is also manifested as a side effect in anticancer mitosis-blocking therapy. The aim of this study was to examine the possibility of preventing side effects of mitostatic drugs in patients with vascular endothelium dysfunction and to establish effective doses able to disrupt the microtubular network without interfering with the endothelial barrier function. Previously, it was found that the population of endothelial cell microtubules is heterogeneous. Along with dynamic microtubules, cell cytoplasm contains a certain amount of post-translationally modified microtubules that are less active and less susceptible to external influences than dynamic microtubules. We have shown that the area occupied with stable microtubules is relatively large (approx. one third of the total cell area). We assume that it can account for a higher resistance of the endothelial monolayer to factors responsible for vascular endothelium dysfunction. This hypothesis was validated in this study, in which nocodazole was used to induce vascular endothelium dysfunction in lung endothelial cells. The effect of nocodazole on endothelial cell cytoskeleton was found to be dose-dependent. Nocodazole in micromolar concentrations not only irreversibly changed the barrier function, but also upset the viability of endothelial cells and induced their death. Nanomolar concentrations of nocodazole also increased the permeability of the endothelial monolayer; this effect was reversible at the drug concentration ranging from 100 to 200 nM. At 100 nM, nocodazole induced partial disruption of the microtubule network near the cell margin without any appreciable effect on acetylated microtubules and actin filaments. At 200 nM, nocodazole exerted a pronounced effect on the system of dynamic (but not acetylated) microtubules and increased the population of actin filaments in the central region of the cell. Our data suggest that disruption of peripheral microtubules triggers a cascade of reactions culminating in endothelial barrier dysfunction; however, the existence of a large population of microtubules resistant to nanomolar concentrations of the drug provides higher viability of endothelial cells and restores their functional activity.

Keywords

Supplement Series Nocodazole Cell Periphery Allicin Dynamic Microtubule 
These keywords were added by machine and not by the authors. This process is experimental and the keywords may be updated as the learning algorithm improves.

References

  1. 1.
    Lum, H. and Malik, A.B., Mechanisms of Increased Endothelial Permeability, Can. J. Physiol. Pharmacol., 1996, vol. 74, pp. 787–800.PubMedCrossRefGoogle Scholar
  2. 2.
    Dudek, S.M. and Garcia, J.G., Cyloskeletal Regulation of Pulmonary Vascular Permeability, J. Appl. Physiol., 2001, vol. 91, pp. 1487–1500.PubMedGoogle Scholar
  3. 3.
    Bogatcheva, N.V., Garcia, J.G.N., and Verin A.D., Molecular Mechanisms of Thrombin-Induced Endothelial Cell Permeability, Biochemistry (Moscow), 2002, vol. 67, no. 1, pp. 75–84.CrossRefGoogle Scholar
  4. 4.
    Birukova, A., Birukov, K., Smurova, K., Kaibuchi, K., Alieva, I., Garcia, J.G., and Verin, A., Novel Role of Microtubules in Thrombin-Induced Endothelial Barrier Dysfunction, FASEB J., 2004, vol. 18, pp. 1879–1890.PubMedCrossRefGoogle Scholar
  5. 5.
    Birukova, A., Smurova, K., Birukov, K., Usatyuk, P., Liu, F., Kaibuchi, K., Ricks-Cord, A., Natarajan, V., Alieva, I., Garcia, J.G., and Verin, A., Microtubule Disassembly Induces Cytoskeletal Remodeling and Vascular Barrier Dysfunction: Role of Rho-Dependent Mechanisms, J. Cell. Physiol., 2004, vol. 201, pp. 55–70.PubMedCrossRefGoogle Scholar
  6. 6.
    Garcia, J.G., Davis, H.W., and Patterson, C.E., Regulation of Endothelial Cell Gap Formation and Barrier Dysfunction: Role of Myosin Light Chain Phosphorylation, J. Cell. Physiol., 1995, vol. 163, pp. 510–522.PubMedCrossRefGoogle Scholar
  7. 7.
    Garcia, J.G., Verin, A.D., and Schaphorst, K.L., Regulation of Thrombin-Mediated Endothelial Cell Contraction and Permeability, Semin. Thromb. Hemostasis, 1996, vol. 22, pp. 309–315.CrossRefGoogle Scholar
  8. 8.
    Van Nieuw Amerongen, G.P., Van Delft S., Vermeer, M.A., Collard, J.G., and van Hinsbergh, V.W., Activation of RhoA by Thrombin in Endothelial Hyperpermeability: Role of Rho Kinase and Protein Tyrosine Kinases, Circ. Res., 2000, vol. 87, pp. 335–340.PubMedGoogle Scholar
  9. 9.
    Groeneveld, A.B., Vascular Pharmacology of Acute Lung Injury and Acute Respiratory Distress Syndrome, Vascul. Pharmacol., 2002, vol. 39, pp. 247–256.PubMedCrossRefGoogle Scholar
  10. 10.
    Smurova, K.M., Birukova, A.A., Garcia, J., Vorobyev, I.A., Alieva, I.B., and Verin, A.D., Thrombin-Induced Rearrangement of the Microtubule System in Lung Endothelium Cells, Tsitologiya (Rus.), 2004, vol. 46, no. 8, pp. 695–703.Google Scholar
  11. 11.
    Smurova, K.M., Birukova, A.A., Verin, A.D., and Alieva, I.B., Microtubule System in Endothelial Barrier Dysfunction: Disassembly of Peripheral Microtubules and Microtubule Reorganization in Internal Cytoplasm, Cell and Tissue Biology, 2008, vol. 2, no. 1, pp. 45–52.CrossRefGoogle Scholar
  12. 12.
    Cattan, C.E. and Oberg, K.C., Vinorelbine Tartrate-Induced Pulmonary Edema Confirmed on Rechallenge, Pharmacotherapy, 1999, vol. 19, no. 8, pp. 992–994.PubMedCrossRefGoogle Scholar
  13. 13.
    Uoshima, N., Yoshioka, K., Tegoshi, H., Wada, S., Fujiwara, Y., Satake, N., Kasamatsu, Y., and Yokoho, S., Acute Respiratory Failure Caused by Vinorelbine Tartrate in a Patient with Non-Small Cell Lung Cancer, Intern. Med., 2001, vol. 40, no. 8, pp. 779–782.PubMedCrossRefGoogle Scholar
  14. 14.
    George, Ph., Journey, L.J., and Goldstein, M., Effect of Vincristine on the Fine Structure of HeLa Cell during Mitosis, J. Natl. Cancer Inst., 1965, vol. 35, pp. 355–365.PubMedGoogle Scholar
  15. 15.
    Krishan, A., Fine Structure of the Kinetochore in Vinblastine Sulphate-Treated Cells, J. Ultrastruct. Res., 1968, vol. 23, pp. 124–143.CrossRefGoogle Scholar
  16. 16.
    Thilagaratnam, C.N. and Main, J.H., Mitostatic Action of Vinblastine Sulphate on Oral Epithelia of Hamsters, J. Oral Pathol., 1972, vol. 1, no. 2, pp. 84–88.PubMedCrossRefGoogle Scholar
  17. 17.
    Zuckerberg, C. and Solari, A.J., Centriolar Changes Induced by Vinblastine Sulphate in the Seminiferous Epithelium of the Mouse, Exp. Cell. Res., 1973, vol. 76, pp. 470–475.PubMedCrossRefGoogle Scholar
  18. 18.
    Alieva, I.B., Gorgidze, L.A., Komarova, Yu.A., Chernobelskaya, O.A., and Vorobjev, I.A., Experimental Model for Studying the Primary Cilia in Tissue Cultured Cells, Membr. Cell. Biol., 1999, vol. 12, no. 6, pp. 895–905.PubMedGoogle Scholar
  19. 19.
    Verin, A.D., Birukova, A., Wang, P., Liu, F., Becker, P., Birukov, K., and Garcia, J.G., Microtubule Disassembly Increases Endothelial Cell Barrier Dysfunction: Role of MLC Phosphorylation, Am. J. Physiol.: Lung Cell Mol. Physiol., 2001, vol. 281, pp. L565–L574.Google Scholar
  20. 20.
    Hirsch, K., Danilenko, M., Giat, J., Miron, T., Rabinkov, A., Wilchek, M., Mirelman, D., Levy, J., and Sharoni, Y., Effect of Purified Allicin, the Major Ingredient of Freshly Crushed Garlic, on Cancer Cell Proliferation, Nutr. Cancer, 2000, vol. 38, no. 2, pp. 245–254.PubMedCrossRefGoogle Scholar
  21. 21.
    Oommen, S., Anto, R.J., Srinivas, G., and Karunagaran, D., Allicin (from Garlic) Induces Caspase-Mediated Apoptosis in Cancer Cells, Eur. J. Pharmacol., 2004, vol. 485, nos. 1–3, pp. 97–103.PubMedCrossRefGoogle Scholar
  22. 22.
    Prager-Khoutorsky, M., Goncharov, I., Rabinkov, A., Mirelman, D., Geiger, B., and Bershadsky, A.D., Allicin Inhibits Cell Polarization, Migration and Division via Its Direct Effect on Microtubules, Cell. Motil. Cytoskeleton, 2007, vol. 64, no. 5, pp. 321–337.PubMedCrossRefGoogle Scholar
  23. 23.
    Aznar, S., Fernandez-Valeron, P., Espina, C., and Lacal, J.C., Rho GTPases: Potential Candidates for Anticancer Therapy, Cancer Lett., 2004, vol. 206, no. 2, pp. 181–191.PubMedCrossRefGoogle Scholar
  24. 24.
    Zhang, B., Rho GDP Dissociation Inhibitors As Potential Targets for Anticancer Treatment, Drug Resist. Updat. 2006, vol. 9, no. 3, pp. 134–141.PubMedCrossRefGoogle Scholar
  25. 25.
    Fritz, G. and Kaina, B., Rho GTPases: Promising Cellular Targets for Novel Anticancer Drugs, Curr. Cancer Drug Targets, 2006, vol. 6, no. 1, pp. 1–14.PubMedCrossRefGoogle Scholar

Copyright information

© MAIK Nauka 2008

Authors and Affiliations

  • K. M. Smurova
    • 1
  • A. A. Birukova
    • 2
  • A. D. Verin
    • 3
  • I. B. Alieva
    • 1
  1. 1.Belozersky Institute of Physico-Chemical BiologyMoscow Lomonosov State UniversityMoscowRussia
  2. 2.Department of Medicine, Section of Pulmonary and Critical Care MedicineUniversity of ChicagoChicagoUSA
  3. 3.Vascular Biology CenterMedical College of GeorgiaAugustaUSA

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