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Originally published as MBC in Press, 10.1091/mbc.E03-09-0646 on December 29, 2003

Vol. 15, Issue 3, 1364-1373, March 2004

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Repression of Na,K-ATPase {beta}1-Subunit by the Transcription Factor Snail in Carcinoma

Cromwell E. Espineda *, Jay H. Chang * {dagger}, Jeffery Twiss {dagger}, Sigrid A. Rajasekaran *, and Ayyappan K. Rajasekaran * {ddagger}

* Department of Pathology and Laboratory Medicine, David Geffen School of Medicine, University of California, Los Angeles, Los Angeles, California 90095; {dagger} Nemours Biomedical Research, Alfred I. duPont Hospital for Children, Wilmington, Delaware 19803

Submitted September 5, 2003; Revised November 17, 2003; Accepted November 29, 2003
Monitoring Editor: Guido Guidotti

The Na,K-ATPase consists of two essential {alpha}- and {beta}-subunits and regulates the intracellular Na+ and K+ homeostasis. Although the {alpha}-subunit contains the catalytic activity, it is not active without functional {beta}-subunit. Here, we report that poorly differentiated carcinoma cell lines derived from colon, breast, kidney, and pancreas show reduced expression of the Na,K-ATPase {beta}1-subunit. Decreased expression of {beta}1-subunit in poorly differentiated carcinoma cell lines correlated with increased expression of the transcription factor Snail known to down-regulate E-cadherin. Ectopic expression of Snail in well-differentiated epithelial cell lines reduced the protein levels of E-cadherin and {beta}1-subunit and induced a mesenchymal phenotype. Reduction of Snail expression in a poorly differentiated carcinoma cell line by RNA interference increased the levels of Na,K-ATPase {beta}1-subunit. Furthermore, Snail binds to a noncanonical E-box in the Na,K-ATPase {beta}1-subunit promoter and suppresses its promoter activity. These results suggest that down-regulation of Na,K-ATPase {beta}1-subunit and E-cadherin by Snail are associated with events leading to epithelial to mesenchymal transition.

Article published online ahead of print. Mol. Biol. Cell 10.1091/mbc.E03-09-0646. Article and publication date are available at www.molbiolcell.org/cgi/doi/10.1091/mbc.E03-09-0646.

Abbreviations used: EMSA, electrophoretic mobility shift assay; EMT, epithelial to mesenchymal transition; MDCK, Madin-Darby Canine kidney; MSV-MDCK, Moloney Sarcoma Virus-transformed MDCK; MRE/GREs, mineralcorticoid/glucocorticoid responsive elements; RNAi, RNA interference; RT-PCR, reverse transcription-polymerase chain reaction.

{ddagger} Corresponding author. E-mail address: arajasekaran{at}mednet.ucla.edu.




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