{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,14]],"date-time":"2026-10-14T23:09:42Z","timestamp":1792019382584,"version":"4.4.0"},"reference-count":45,"publisher":"Wiley","issue":"4","license":[{"start":{"date-parts":[[2020,1,6]],"date-time":"2020-01-06T00:00:00Z","timestamp":1578268800000},"content-version":"vor","delay-in-days":0,"URL":"http:\/\/onlinelibrary.wiley.com\/termsAndConditions#vor"}],"funder":[{"name":"Ministry of Sciences and Technology in Taiwan","award":["MOST 107-2320-B-002-018-MY3"],"award-info":[{"award-number":["MOST 107-2320-B-002-018-MY3"]}]},{"name":"Ministry of Sciences and Technology in Taiwan","award":["MOST 106-2320-B-002-005-MY3"],"award-info":[{"award-number":["MOST 106-2320-B-002-005-MY3"]}]}],"content-domain":{"domain":["onlinelibrary.wiley.com"],"crossmark-restriction":true},"short-container-title":["The Prostate"],"published-print":{"date-parts":[[2020,3]]},"abstract":"<jats:title>Abstract<\/jats:title><jats:sec><jats:title>Background<\/jats:title><jats:p>Cardiac glycosides, which inhibit Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase, display inotropic effects for the treatment of congestive heart failure and cardiac arrhythmia. Recent studies have suggested signaling downstream of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase action in the regulation of cell proliferation and apoptosis and have revealed the anticancer activity of cardiac glycosides. The study aims to characterize the anticancer potential of ascleposide, a natural cardenolide, and to uncover its primary target and underlying mechanism against human castration\u2010resistant prostate cancer (CRPC).<\/jats:p><\/jats:sec><jats:sec><jats:title>Methods<\/jats:title><jats:p>Cell proliferation was examined in CRPC PC\u20103 and DU\u2010145 cells using sulforhodamine B assay, carboxyfluorescein succinimidyl ester staining assay and clonogenic examination. Flow cytometric analysis was used to detect the distribution of cell cycle phase, mitochondrial membrane potential, intracellular Na<jats:sup>+<\/jats:sup> and Ca<jats:sup>2+<\/jats:sup> levels, and reactive oxygen species production. Protein expression was examined using Western blot analysis. Endocytosis of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase was determined using <jats:italic>confocal immunofluorescence<\/jats:italic> microscopic <jats:italic>examination<\/jats:italic>.<\/jats:p><\/jats:sec><jats:sec><jats:title>Results<\/jats:title><jats:p>Ascleposide induced an increase of intracellular Na<jats:sup>+<\/jats:sup> and a potent antiproliferative effect. It also induced a decrease of G1 phase distribution while an increase in both G2\/M and apoptotic sub\u2010G1 phases, and downregulated several cell cycle regulator proteins, including cyclins, Cdk, p21, and p27 Cip\/Kip proteins, Rb and c\u2010Myc. Ascleposide decreased the expression of antiapoptotic Bcl\u20102 members (eg, Bcl\u20102 and Mcl\u20101) but upregulated proapoptotic member (eg, Bak), leading to a significant loss of mitochondrial membrane potential and activation of both caspase\u20109 and caspase\u20103. Ascleposide also dramatically induced tubulin acetylation, leading to inhibition of the catalytic activity of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase. Notably, extracellular high K<jats:sup>+<\/jats:sup> (16\u2009mM) significantly blunted ascleposide\u2010mediated effects. Furthermore, ascleposide induced a p38 MAPK\u2010dependent endocytosis of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase and downregulated the protein expression of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase \u03b11 subunit.<\/jats:p><\/jats:sec><jats:sec><jats:title>Conclusion<\/jats:title><jats:p>Ascleposide displays antiproliferative and apoptotic activities dependent on the inhibition of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase pumping activity through p38 MAPK\u2010mediated endocytosis of Na<jats:sup>+<\/jats:sup>\/K<jats:sup>+<\/jats:sup>\u2010ATPase and downregulation of \u03b11 subunit, which in turn cause tubulin acetylation and cell cycle arrest. Cell apoptosis is ultimately triggered by the activation of caspase cascade attributed to mitochondrial damage through the downregulation of Bcl\u20102 and Mcl\u20101 protein expressions while upregulation of Bak protein levels. The data also suggest the potential of ascleposide in anti\u2010CRPC development.<\/jats:p><\/jats:sec>","DOI":"10.1002\/pros.23944","type":"journal-article","created":{"date-parts":[[2020,1,6]],"date-time":"2020-01-06T17:59:39Z","timestamp":1578333579000},"page":"305-318","update-policy":"https:\/\/doi.org\/10.1002\/crossmark_policy","source":"Crossref","is-referenced-by-count":10,"title":["Ascleposide, a natural cardenolide, induces anticancer signaling in human castration\u2010resistant prostatic cancer through Na<sup>+<\/sup>\/K<sup>+<\/sup>\u2010ATPase internalization and tubulin acetylation"],"prefix":"10.1002","volume":"80","author":[{"given":"Wohn\u2010Jenn","family":"Leu","sequence":"first","affiliation":[{"name":"School of Pharmacy National Taiwan University Taipei Taiwan"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Ching\u2010Ting","family":"Wang","sequence":"additional","affiliation":[{"name":"School of Pharmacy National Taiwan University Taipei 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