Vacuolar ATPase driven potassium transport in highly metastatic breast cancer cells

Sarah A. Salyer, Jordan R. Olberding, Anthony A. Distler, Eleanor D. Lederer, Barbara J. Clark, Nicholas A Delamere, Syed J. Khundmiri

Research output: Contribution to journalArticle

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Abstract

Breast cancer is the second leading cause of death in women and thus has received a great deal of attention by researchers. Recent studies suggested decreased occurrence of cancer in patients treated with cardiac glycosides (CGs) for heart conditions. Because CGs induce their cellular effects via the Na+, K+ ATPase (Na-K), we treated four breast cancer cell lines (MCF-7, T47D, MDA-MB453, and MDA-MB231) and a non-cancerous breast ductal epithelial cell line (MCF-10A) with ouabain, a well-characterized CG, and measured cell proliferation by measuring bromodeoxyuridine incorporation. Ouabain (1μM) decreased cell proliferation in all cell lines studied except MDA-MB453 cells. Western blot of Na-K α and β subunits showed α1, α3, and β1 expression in all cell lines except MDA-MB453 cells where Na-K protein and mRNA were absent. Potassium uptake, measured as rubidium (86Rb) flux, and intracellular potassium were both significantly higher in MDA-MB453 cells compared to MCF-10A cells. RT-qPCR suggested a 7 fold increase in voltage-gated potassium channel (KCNQ2) expression in MDA-MB453 cells compared to MCF-10A cells. Inhibition of KCNQ2 prevented cell growth and 86Rb uptake in MDA-MB453 cells but not in MCF-10A cells. All cancer cells had significantly higher vacuolar H-ATPase (V-ATPase) activity than MCF-10A cells. Inhibition of V-ATPase decreased 86Rb uptake and intracellular potassium in MDA-MB453 cells but not in MCF-10A cells. The findings point to the absence of Na-K, high hERG and KCNQ2 expression, elevated V-ATPase activity and sensitivity to V-ATPase inhibitors in MDA-MB453. We conclude that cancer cells exhibit fundamentally different metabolic pathways for maintenance of intracellular ion homeostasis.

Original languageEnglish (US)
Pages (from-to)1734-1743
Number of pages10
JournalBiochimica et Biophysica Acta - Molecular Basis of Disease
Volume1832
Issue number10
DOIs
StatePublished - Oct 2013

Fingerprint

Vacuolar Proton-Translocating ATPases
Potassium
Breast Neoplasms
Cardiac Glycosides
Cell Line
Ouabain
Cell Proliferation
Rubidium
Voltage-Gated Potassium Channels
Neoplasms
Bromodeoxyuridine
Metabolic Networks and Pathways

Keywords

  • Breast cancer cell
  • MDA-MB453 cell
  • Na-K ATPase
  • Ouabain
  • Potassium transport
  • Vacuolar H-ATPase

ASJC Scopus subject areas

  • Molecular Biology
  • Molecular Medicine

Cite this

Vacuolar ATPase driven potassium transport in highly metastatic breast cancer cells. / Salyer, Sarah A.; Olberding, Jordan R.; Distler, Anthony A.; Lederer, Eleanor D.; Clark, Barbara J.; Delamere, Nicholas A; Khundmiri, Syed J.

In: Biochimica et Biophysica Acta - Molecular Basis of Disease, Vol. 1832, No. 10, 10.2013, p. 1734-1743.

Research output: Contribution to journalArticle

Salyer, Sarah A. ; Olberding, Jordan R. ; Distler, Anthony A. ; Lederer, Eleanor D. ; Clark, Barbara J. ; Delamere, Nicholas A ; Khundmiri, Syed J. / Vacuolar ATPase driven potassium transport in highly metastatic breast cancer cells. In: Biochimica et Biophysica Acta - Molecular Basis of Disease. 2013 ; Vol. 1832, No. 10. pp. 1734-1743.
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AB - Breast cancer is the second leading cause of death in women and thus has received a great deal of attention by researchers. Recent studies suggested decreased occurrence of cancer in patients treated with cardiac glycosides (CGs) for heart conditions. Because CGs induce their cellular effects via the Na+, K+ ATPase (Na-K), we treated four breast cancer cell lines (MCF-7, T47D, MDA-MB453, and MDA-MB231) and a non-cancerous breast ductal epithelial cell line (MCF-10A) with ouabain, a well-characterized CG, and measured cell proliferation by measuring bromodeoxyuridine incorporation. Ouabain (1μM) decreased cell proliferation in all cell lines studied except MDA-MB453 cells. Western blot of Na-K α and β subunits showed α1, α3, and β1 expression in all cell lines except MDA-MB453 cells where Na-K protein and mRNA were absent. Potassium uptake, measured as rubidium (86Rb) flux, and intracellular potassium were both significantly higher in MDA-MB453 cells compared to MCF-10A cells. RT-qPCR suggested a 7 fold increase in voltage-gated potassium channel (KCNQ2) expression in MDA-MB453 cells compared to MCF-10A cells. Inhibition of KCNQ2 prevented cell growth and 86Rb uptake in MDA-MB453 cells but not in MCF-10A cells. All cancer cells had significantly higher vacuolar H-ATPase (V-ATPase) activity than MCF-10A cells. Inhibition of V-ATPase decreased 86Rb uptake and intracellular potassium in MDA-MB453 cells but not in MCF-10A cells. The findings point to the absence of Na-K, high hERG and KCNQ2 expression, elevated V-ATPase activity and sensitivity to V-ATPase inhibitors in MDA-MB453. We conclude that cancer cells exhibit fundamentally different metabolic pathways for maintenance of intracellular ion homeostasis.

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