title: Ouabain targets the unfolded protein response for selective killing of HepG2 cells during glucose deprivation.
abstract: Ouabain is a cardiotonic steroid and specific inhibitor of the Na(+)/K(+)-ATPase. The relationship between ouabain treatment and the unfolded protein response (UPR) in cells is not precisely understood. Therefore, we studied the possible effects of ouabain on proliferation, apoptosis, and the UPR. HepG2 cells were cultured overnight and then treated with various concentrations of ouabain (0.75 to 750 nM) in the absence or presence of 10 mM 2-deoxyglucose (2-DG) for 48 hours. We also used real-time polymerase chain reaction to obtain quantitative measurements of expression levels of Grp78, Grp94, CHOP, MTJ-1, HKII, MDR-1, MRP-1, HO-1, and Par-4. Cell number, viability, and proliferation of HepG2 cells were monitored with a real-time cell analyzer system (xCELLigence). We show that ouabain modulates the UPR transcription program and induces cell death in glucose-deprived tumor cells. Ouabain at all concentrations showed no cytotoxicity whereas all concentrations were very effective under 2-DG stress conditions. Our findings show that disruption of the UPR during glucose deprivation could be an attractive approach for selective cancer cell killing and could provide a chemical basis for developing UPR-targeting drugs against solid tumors. Ouabain use as an adjunct to conventional cancer therapy also warrants vigorous investigation.
authors: Gazi State Hospital, Samsun, Turkey.
authors: Tulay
authors: T
authors: Ozdemir
authors: Rukiye
authors: R
authors: Nar
authors: Veli
authors: V
authors: Kilinc
authors: Hasan
authors: H
authors: Alacam
authors: Osman
authors: O
authors: Salis
authors: Aynur
authors: A
authors: Duzgun
authors: Sedat
authors: S
authors: Gulten
authors: Abdulkerim
authors: A
authors: Bedir
mesh_terms: D000818:Animals; D017209:Apoptosis / Q000187:drug effects; D048708:Cell Growth Processes / Q000187:drug effects; D003847:Deoxyglucose / Q000494:pharmacology; D000091342:Endoplasmic Reticulum Chaperone BiP; D015870:Gene Expression; D005947:Glucose / Q000172:deficiency* / Q000378:metabolism; D050956:HSP40 Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D006360:Heat-Shock Proteins / Q000096:biosynthesis / Q000235:genetics; D051547:Heme Oxygenase-1 / Q000096:biosynthesis / Q000235:genetics; D056945:Hep G2 Cells; D006801:Humans; D008562:Membrane Glycoproteins / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D009363:Neoplasm Proteins / Q000096:biosynthesis / Q000235:genetics; D010042:Ouabain / Q000494:pharmacology*; D060888:Real-Time Polymerase Chain Reaction; D015398:Signal Transduction; D056811:Unfolded Protein Response / Q000187:drug effects* / Q000235:genetics
chemical_list: C092319:Dnajc1 protein, mouse; D000091342:Endoplasmic Reticulum Chaperone BiP; D050956:HSP40 Heat-Shock Proteins; C000717247:HSPA5 protein, human; D006360:Heat-Shock Proteins; C000717248:Hspa5 protein, mouse; D008562:Membrane Glycoproteins; D009363:Neoplasm Proteins; C053387:endoplasmin; D010042:Ouabain; D003847:Deoxyglucose; D051547:Heme Oxygenase-1; D005947:Glucose