21-Benzylidene Digoxin: A Proapoptotic Cardenolide of Cancer Cells That Up-Regulates Na,K-ATPase and Epithelial Tight Junctions

Abstract

Cardiotonic steroids are used to treat heart failure and arrhythmia and have promising anticancer effects. The prototypic cardiotonic steroid ouabain may also be a hormone that modulates epithelial cell adhesion. Cardiotonic steroids consist of a steroid nucleus and a lactone ring, and their biological effects depend on the binding to their receptor, Na,K-ATPase, through which, they inhibit Na+ and K+ ion transport and activate of several intracellular signaling pathways. In this study, we added a styrene group to the lactone ring of the cardiotonic steroid digoxin, to obtain 21-benzylidene digoxin (21-BD), and investigated the effects of this synthetic cardiotonic steroid in different cell models. Molecular modeling indicates that 21-BD binds to its target Na,K-ATPase with low affinity, adopting a different pharmacophoric conformation when bound to its receptor than digoxin. Accordingly, 21-DB, at relatively high µM amounts inhibits the activity of Na,K-ATPase α1, but not α2 and α3 isoforms. In addition, 21-BD targets other proteins outside the Na,K-ATPase, inhibiting the multidrug exporter Pdr5p. When used on whole cells at low µM concentrations, 21-BD produces several effects, including: 1) up-regulation of Na,K-ATPase expression and activity in HeLa and RKO cancer cells, which is not found for digoxin, 2) cell specific changes in cell viability, reducing it in HeLa and RKO cancer cells, but increasing it in normal epithelial MDCK cells, which is different from the response to digoxin, and 3) changes in cell-cell interaction, altering the molecular composition of tight junctions and elevating transepithelial electrical resistance of MDCK monolayers, an effect previously found for ouabain. These results indicate that modification of the lactone ring of digoxin provides new properties to the compound, and shows that the structural change introduced could be used for the design of cardiotonic steroid with novel functions.

DOI: 10.1371/journal.pone.0108776

Extracted Key Phrases

8 Figures and Tables

Statistics

01000200020162017
Citations per Year

372 Citations

Semantic Scholar estimates that this publication has 372 citations based on the available data.

See our FAQ for additional information.

Cite this paper

@inproceedings{Rocha201421BenzylideneDA, title={21-Benzylidene Digoxin: A Proapoptotic Cardenolide of Cancer Cells That Up-Regulates Na,K-ATPase and Epithelial Tight Junctions}, author={Sayonarah C. Rocha and Marco Tulio C Pessoa and Luiza D. R. Neves and Silmara L. G. Alves and Luciana Maria Silva and H{\'e}rica de Lima Santos and Soraya M. F. Oliveira and Alex G. Taranto and Moacyr Comar and Isabella V. Gomes and F{\'a}bio Vieira dos Santos and Natasha Paix{\~a}o and Luis Eduardo Menezes Quintas and François G No{\"{e}l and Ant{\'o}nio Nazar{\'e} Pereira and Ana C. S. C. Tessis and Nat{\'a}lia Gomes and Otac{\'i}lio da Cruz Moreira and Ruth Rincon-Heredia and Fernando de Pilla Varotti and Gustavo Blanco and Jos{\'e} Augusto Ferreira Perez Villar and Ruben Contreras and Leandro A Barbosa and Shree R Singh}, booktitle={PloS one}, year={2014} }