title: Amiloride kills malignant glioma cells independent of its inhibition of the sodium-hydrogen exchanger.
abstract: Previously, we demonstrated that malignant glioma cell lines have increased intracellular pH (pHi) as a result of increased activities of the type I sodium/hydrogen exchanger (NHE1). This alkalotic pHi of 7.2 to 7.4 is favorable for augmented glycolysis, DNA synthesis, and cell cycle progression. Conversely, reductions in pHi have been associated with reduced rates of proliferation in transformed cell types. The effects of reducing pHi directly and by NHE1 inhibition on human malignant glioma cells were systematically compared with those on primary rat astrocytes. Neither cariporide, nor direct acidification to pHi 6.9 altered the proliferative rates or viabilities of human U87 or U118 malignant glioma cell lines. However, amiloride significantly impaired glioma cell proliferation and viability while not affecting astrocytes at concentrations (500 microM) that exceeded its inhibition of NHE1 in glioma cells (IC50 = 17 microM). Preventing a reduction of pHi did not alter the drug's antiproliferative and cytotoxic effects on glioma cells. These findings indicated that amiloride's cytotoxic effects on glioma cells are independent of its ability to inhibit NHE1 or to reduce intracellular pHi. The amiloride derivative 2,4 dichlorobenzamil (DCB) inhibits the sodium-calcium exchanger (NCX) and was both antiproliferative and cytotoxic to glioma cells at low doses (20 microM). By contrast, KB-R7943 [(2-[2-[4-nitrobenzyloxy]phenyl]ethyl)-isothioureamethanesulfonate] preferentially blocks sodium-dependent calcium influx by NCX (reverse mode) and was nontoxic to glioma cells. It is proposed that DCB (20 microM) and amiloride (500 microM) impair calcium efflux by NCX, leading to elevations of intracellular calcium that initiate a morphologically necrotic, predominantly caspase-independent glioma cell death.
authors.affiliation: Department of Neurology, School of Medicine, University of California Davis, Davis, California 95616, USA.
authors.forename: Manu
authors.initials: M
authors.lastname: Hegde
authors.forename: Jane
authors.initials: J
authors.lastname: Roscoe
authors.forename: Peter
authors.initials: P
authors.lastname: Cala
authors.forename: Fredric
authors.initials: F
authors.lastname: Gorin
mesh_terms: D000584:Amiloride / Q000031:analogs & derivatives* / Q000494:pharmacology*; D000818:Animals; D001253:Astrocytes / Q000166:cytology / Q000187:drug effects; D020169:Caspases / Q000378:metabolism; D002455:Cell Division / Q000187:drug effects; D002470:Cell Survival / Q000187:drug effects; D004789:Enzyme Activation; D005910:Glioma / Q000473:pathology*; D006801:Humans; D006863:Hydrogen-Ion Concentration; D051381:Rats; D019831:Sodium-Calcium Exchanger / Q000037:antagonists & inhibitors; D017923:Sodium-Hydrogen Exchangers / Q000037:antagonists & inhibitors*; D014407:Tumor Cells, Cultured
chemical_list: D019831:Sodium-Calcium Exchanger; D017923:Sodium-Hydrogen Exchangers; C041470:3',4'-dichlorobenzamil; D000584:Amiloride; D020169:Caspases