Inhibition of 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) endocytosis by ouabain in human endothelial cells

FEBS Letters
Lucia TrevisiSisto Luciani

Abstract

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT) uptake and reduction is widely used to evaluate cell proliferation and viability. MTT is taken up by the cells through endocytosis. We find that ouabain (1-200 nM) inhibits MTT reduction in human umbilical vein endothelial cells (HUVEC) without affecting cell viability. Ouabain does not inhibit MTT reduction when cell lysates substituted for the intact cells. Disruption of caveolae by cholesterol depletion, completely prevents the effect of ouabain. Treatment of HUVEC with Src inhibitor 4-amino-5-(4-chlorophenyl)-7-(t-butyl)pyrazolo[3,4-d]pyrimidine partially abrogates the inhibitory effect of ouabain. The data suggest that ouabain interaction with caveolar Na/K-ATPase inhibits MTT endocytosis through the activation of signaling proteins such as Src kinase.

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Citations

Oct 17, 2008·Photomedicine and Laser Surgery·Aline Helena Araujo MachadoNewton Soares Da Silva
Mar 3, 2007·The Journal of Pharmacy and Pharmacology·Chang Hwa JungHo Moon Seog
Mar 10, 2012·Toxicology in Vitro : an International Journal Published in Association with BIBRA·Lanhai LüJie Xu
Apr 20, 2017·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·Sergei N OrlovOlga D Lopina
Aug 14, 2012·Molecules : a Journal of Synthetic Chemistry and Natural Product Chemistry·João Carlos Lima Rodrigues PitaMarianna Vieira Sobral Castello-Branco
Jul 4, 2015·British Journal of Pharmacology·Luigia Di FrancescoPaola Patrignani
Jun 14, 2013·American Journal of Physiology. Renal Physiology·Gustavo Blanco, Darren P Wallace

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