Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells
Abstract The Na+/K+-ATPase (NKA) complex is the master regulator of membrane potential and a target for anti-cancer therapies. Here, we investigate the effect of drug-induced oxidative stress on NKA activity. The natural product, plumbagin increases oxygen radicals through inhibition of oxidative ph...
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2019-08-01
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doaj-711cb0ad0da842b3aed61841711db73e2020-12-08T09:51:48ZengNature Publishing GroupScientific Reports2045-23222019-08-019111010.1038/s41598-019-47261-xPlumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cellsYousef Alharbi0Arvinder Kapur1Mildred Felder2Lisa Barroilhet3Timothy Stein4Bikash R. Pattnaik5Manish S. Patankar6Department of Obstetrics and Gynecology, University of Wisconsin-MadisonDepartment of Obstetrics and Gynecology, University of Wisconsin-MadisonDepartment of Obstetrics and Gynecology, University of Wisconsin-MadisonDepartment of Obstetrics and Gynecology, University of Wisconsin-MadisonDepartment of Comparative Biosciences, School of Veterinary Medicine, University of Wisconsin-MadisonDepartment of Pediatrics and Ophthalmology and Visual Sciences, University of Wisconsin-MadisonDepartment of Obstetrics and Gynecology, University of Wisconsin-MadisonAbstract The Na+/K+-ATPase (NKA) complex is the master regulator of membrane potential and a target for anti-cancer therapies. Here, we investigate the effect of drug-induced oxidative stress on NKA activity. The natural product, plumbagin increases oxygen radicals through inhibition of oxidative phosphorylation. As a result, plumbagin treatment results in decreased production of ATP and a rapid increase in intracellular oxygen radicals. We show that plumbagin induces apoptosis in canine cancer cells via oxidative stress. We use this model to test the effect of oxidative stress on NKA activity. Using whole-cell patch-clamp electrophysiology we demonstrate that short-term exposure (4 min) to plumbagin results in 48% decrease in outward current at +50 mV. Even when exogenous ATP was supplied to the cells, plumbagin treatment resulted in 46% inhibition of outward current through NKA at +50 mV. In contrast, when the canine cancer cells were pre-treated with the oxygen radical scavenger, N-acetylcysteine, the NKA inhibitory activity of plumbagin was abrogated. These experiments demonstrate that the oxidative stress-causing agents such as plumbagin and its analogues, are a novel avenue to regulate NKA activity in tumors.https://doi.org/10.1038/s41598-019-47261-x |
collection |
DOAJ |
language |
English |
format |
Article |
sources |
DOAJ |
author |
Yousef Alharbi Arvinder Kapur Mildred Felder Lisa Barroilhet Timothy Stein Bikash R. Pattnaik Manish S. Patankar |
spellingShingle |
Yousef Alharbi Arvinder Kapur Mildred Felder Lisa Barroilhet Timothy Stein Bikash R. Pattnaik Manish S. Patankar Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells Scientific Reports |
author_facet |
Yousef Alharbi Arvinder Kapur Mildred Felder Lisa Barroilhet Timothy Stein Bikash R. Pattnaik Manish S. Patankar |
author_sort |
Yousef Alharbi |
title |
Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells |
title_short |
Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells |
title_full |
Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells |
title_fullStr |
Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells |
title_full_unstemmed |
Plumbagin-induced oxidative stress leads to inhibition of Na+/K+-ATPase (NKA) in canine cancer cells |
title_sort |
plumbagin-induced oxidative stress leads to inhibition of na+/k+-atpase (nka) in canine cancer cells |
publisher |
Nature Publishing Group |
series |
Scientific Reports |
issn |
2045-2322 |
publishDate |
2019-08-01 |
description |
Abstract The Na+/K+-ATPase (NKA) complex is the master regulator of membrane potential and a target for anti-cancer therapies. Here, we investigate the effect of drug-induced oxidative stress on NKA activity. The natural product, plumbagin increases oxygen radicals through inhibition of oxidative phosphorylation. As a result, plumbagin treatment results in decreased production of ATP and a rapid increase in intracellular oxygen radicals. We show that plumbagin induces apoptosis in canine cancer cells via oxidative stress. We use this model to test the effect of oxidative stress on NKA activity. Using whole-cell patch-clamp electrophysiology we demonstrate that short-term exposure (4 min) to plumbagin results in 48% decrease in outward current at +50 mV. Even when exogenous ATP was supplied to the cells, plumbagin treatment resulted in 46% inhibition of outward current through NKA at +50 mV. In contrast, when the canine cancer cells were pre-treated with the oxygen radical scavenger, N-acetylcysteine, the NKA inhibitory activity of plumbagin was abrogated. These experiments demonstrate that the oxidative stress-causing agents such as plumbagin and its analogues, are a novel avenue to regulate NKA activity in tumors. |
url |
https://doi.org/10.1038/s41598-019-47261-x |
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