We report for the first time a potent apoptotic effect of omeprazole (OM). Apoptosis was induced in Jurkat cells in a time and concentration-dependent mode. Caspase 3 and PARP were rapidly cleaved in response to OM, but apoptosis was only partially inhibited by the caspase 3 inhibitor DEVD-CHO. OM also induced an early lysosomal destabilization which increased progressively and was correlated with a parallel increase in apoptotic cells. The cysteine protease inhibitor E64d gave strong protection against apoptosis thus proving the involvement of lysosomal enzymes in OM-induced apoptosis whereas, it did not impede the caspase 3 cleavage. Instead ZVAD-fmk, a general caspase inhibitor, also able to inhibit cathepsin activity, protected cells completely from OM-induced apoptosis. It therefore seems that both caspases and cysteine cathepsins are involved in the execution stage of OM-induced apoptosis.
WallmarkB.1986. Mechanism of action of omeprazole. Scand. J. Gastroenterol.118(S): 11.
2.
LindbergP.NordbergP.AlmingerT.BrändströmA.WallmarkB., 1986. The mechanism of action of the gastric acid secretion inhibitor omeprazole. J. Med. Chem.29:1327.
3.
WallmarkB.LindbergP., 1987. Mechanism of action of omeprazole. ISI Atlas Science Pharmacol. 1:158.
4.
WallmarkB.BrändströmA.LarssonH., 1984. Evidence for acid-induced transformation of omeprazole into an active inhibitor of (H+ + K+)-ATPase within the parietal cell. Biochim. Biophys. Acta.778:549.
5.
MattssonJ.P.VäänänenK.WallmarkB.LorentzonP., 1991. Omeprazole and bafilomycin, two proton pump inhibitors: Differentiation of their effects on gastric, kidney and bone H(+)-translocating ATPases. Biochim. Biophys. Acta1065: 261.
6.
YamamotoO.TanakaH.UedaF.KimuraK., 1989. Inhibition of H+,K+-ATPase by methyl(E)-2-(3,4-dimethoxystyryl)-benzimidazole-4-carboxylate(ALE-36). Scand. J. Gastroenterol.162(S):178.
7.
SuzukiM.MoriM.MiuraS.SuematsuM.FukumuraD.KimuraH.IshiiH., 1996. Omeprazole attenuates oxygen-derived free radical production from human neutrophils. Free Radic. Biol. Med.21:727.
8.
SabolicI.BrownD.VerbavatzJ.M.KleinmanJ., 1994. H(+)-ATPases of renal cortical and medullary endosomes are differentially sensitive to Sch-28080 and omeprazole. Am. J. Physiol.266: F868.
9.
ForgacM.1989. Structure and function of vacuolar class of ATP-driven proton pumps. Physiol. Rev.69:765.
10.
RudnickG.1986. ATP-driven H+ pumping into intracellular organelles. Annu. Rev. Physiol.48:403.
11.
MellmannI.FuchsR.HeleniusA., 1986. Acidification of the endocytic and exocytic pathways. Annu. Rev. Biochem.55:663.
12.
ClagueM.J.UrbeS.AnientoF.GruenbergJ., 1994. Vacuolar ATPase activity is required for endosomal carrier vesicle formation. J. Biol. Chem.269:21.
13.
ReavesB.BantingG., 1994. Vacuolar ATPase inactivation blocks recycling to the trans-Golgi network from the plasma membrane. FEBS Lett. 345:61.
14.
ManabeT.YoshimoriT.HenomatsuN.TashiroY., 1993. Inhibitors of vacuolar-type H(+)-ATPase suppresses proliferation of cultured cells. J. Cell. Physiol.157:445.
15.
SaurinA.J.HamlettJ.ClagueM.J.PenningtonS.R., 1996. Inhibition of mitogen-induced DNA synthesis by bafilomycin A1 in Swiss 3T3 fibroblasts. Biochem. J.313:65.
16.
HeinleS.StunkelK.ZahnerH.DrautzH.BesslerW.G., 1988. Immunosuppressive effects of the macrolide antibiotic bafilomycin towards lymphocytes and lymphoid cell lines. Arzneimittelforschung38: 1130.
17.
KingA.C.Hernaez-DavisL.CuatrecasasP., 1981. Lysosomotropic amines inhibit mitogenesis induced by growth factors. Proc. Natl. Acad. Sci. USA.78:717.
18.
NishiharaT.AkifusaS.KosekiT.KatoS.MuroM.HanadaN., 1995. Specific inhibitors of vacuolar type H(+)-ATPases induce apoptotic cell death. Biochem. Biophys. Res. Commun.212:255.
19.
AkifusaS.OhguchiM.KosekiT.NaraK.SembaI.YamatoK.OkahashiN.MerinoR.NúñezG.HanadaN.TakeharaT.NishiharaT., 1998. Increase in Bcl-2 level promoted by CD40 ligation correlates with inhibition of B cell apoptosis induced by vacuolar type H(+)-ATPase inhibitor. Exp. Cell. Res.238:82.
20.
TanigakiK.SasakiS.OhkumaS., 2003. In bafilomycin A1-resistant cells, bafilomycin A1 raised lysosomal pH and both prodigiosins and concanamycin A inhibited growth through apoptosis. FEBS Lett. 537:79.
21.
AntunesF.CadenasE.BrunkU.T., 2001. Apoptosis induced by exposure to a low steady-state concentration of H2O2 is a consequence of lysosomal rupture. Biochem. J.356:549.
22.
BrunkU.T.DalenH.RobergK.HellquistH.B., 1997. Photo-oxidative disruption of lysosomal membranes causes apoptosis of cultured human fibroblasts. Free Radic. Biol. Med.23:616.
23.
LiW.YuanX.NordgrenG.DalenH.DubowchikG.M.FirestoneR.A.BrunkU.T., 2000. Induction of cell death by the lysosomotropic detergent MSDH. FEBS Lett. 470:35.
24.
KågedalK.ZhaoM.SvenssonI.BrunkU.T., 2001. Sphingosine-induced apoptosis is dependent on lysosomal proteases. Biochem. J.359:335.
25.
NeuzilJ.SvenssonI.WeberT.WeberC.BrunkU.T., 1999. Alpha-tocopheryl succinate-induced apoptosis in Jurkat T cells involves caspase-3 activation, and both lysosomal and mitochondrial destabilisation. FEBS Lett. 445:295.
26.
HishitaT.Tada-OikawaS.TohyamaK.MiuraY.NishiharaT.TohyamaY.YoshidaY.UchiyamaT.KawanishS., 2001. Caspase-3 activation by lysosomal enzymes in cytochrome c-independent apoptosis in myelodysplastic syndrome-derived cell line P39. Cancer Res. 61:2878.
27.
CacoubP.SbaiA.Costedoat-ChalumeauN.BellangerJ.GodeauP.PietteJ.C., 2000. Edematous syndrome revealing an exudative lymphocytic gastritis: Efficacy of omeprazole. Gastroenterol. Clin. Biol.10:963.
28.
LohoffM.RollinghoffM.SommerF., 2000. Helicobacter pylori gastritis: A Th1 mediated diseas?J. Biotechnol.83:33.
29.
D'EliosM. M.AmedeiA.Del PreteG., 2003. Helicobacter pylori antigen-specific T-cell responses at gastric level in chronic gastritis, peptic ulcer, gastric cancer and low-grade mucosa-associated lymphoid tissue (MALT) lymphoma. Microbes Infect. 5:723.
30.
GuindiM.2000. Role of activated host cells in the promotion of MALT lymphoma growth. Semin. Cancer Biol. 10:341.
31.
RenZ.PangG.ClancyR.LiL.C.LeeC.S.BateyR.BorodyT.DunkleyM., 2001. Shift of the gastric T-cell response in gastric carcinoma. J. Gastroenterol. Hepatol.16:142.
32.
RiedelS.KraftM.KucharzikT.PauelsH.G.TiemannM.SteinbuchelA.DomschkeW.LugeringN., 2001. CD4+ Th1-cells predominate in low-grade B-cell limphoma of gastric mucosa-associated lymphoid tissue (MALT type). Scand. J. Gastroenterol.36:1198.
33.
NicolettiI.MiglioratiG.PagliacciM.C.GrignaniF.RiccardiC., 1991. A rapid and simple method for measuring thymocyte apoptosis by propidium iodide staining and flow cytometry. J. Immunol. Methods139: 271.
34.
NeuzilJ.WeberT.SchröderA.LuM.OstermannG.GellertN.MayneG.C.OlejnickaB.Nègre-SalvayreA.StichaM.CoffeyR.J.WeberC., 2001. Induction of cancer cell apoptosis by alpha-tocopheryl succinate: Molecular pathways and structural requirements. FASEB J. 15:403.
35.
RobergK.2001. Relocalization of cathepsin D and cytochrome c early in apoptosis revealed by immunoelectron microscopy. Lab. Invest.81: 149.
36.
RobergK.ÖllingerK., 1998. Oxidative stress causes relocation of the lysosomal enzyme cathepsin D with ensuing apoptosis in neonatal rat cardiomyocytes. Am. J. Pathol.152:1151.
37.
OhsawaY.IsaharaK.KanamoriS.ShibataM.KametakaS.GotowT.WatanabeT.KominamiE.UchiyamaY., 1998. An ultrastructural and immunohistochemical study of PC12 cells during apoptosis induced by serum deprivation with special reference to autophagy and lysosomal cathepsins. Arch. Histol. Cytol.61:395.
38.
IsaharaK.OhsawaY.KanamoriS.ShibataM.WaguriS.SatoN.GotowT.WatanabeT.MomoiT.UraseK.KominamiE.UchiyamaY., 1999. Regulation of a novel pathway for cell death by lysosomal aspartic and cysteine proteinases. Neuroscience91: 233.
39.
JhonsonD.E.2000. Noncaspase proteases in apoptosis. Leukemia14: 1695.
40.
GuicciardiM.E.DeussingJ.MiyoshiH.BronkS.F.SvingenP.A.PetersC.KaufmannS.H.GoresG.J., 2000. Cathepsin B contributes to TNF-alpha-mediated hepatocyte apoptosis by promoting mitochondrial release of cytochrome c. J. Clin. Invest.106:1127.
41.
FoghsgaardL.WissingD.MauchD.LademannU.BastholmL.BoesM.EllingF.LeistM.JäätteläM., 2001. Cathepsin B acts as a dominant execution protease in tumor cell apoptosis induced by tumor necrosis factor. J. Cell. Biol.153:999.
42.
TakumaK.KiriuM.MoriK.LeeE.EnomotoR.BabaA.MatsudaT., 2003. Roles of cathepsins in reperfusion-induced apoptosis in cultured astrocytes. Neurochem. Int.42:153.
43.
BiswasK.BandyopadhyayU.ChattopadhyayI.VaradarajA.AliE.BanerjeeR.K., 2003. A novel antioxidant and antiapoptotic role of omeprazole to block gastric ulcer through scavenging of hydroxyl radical. J. Biol. Chem.278:10993.
44.
AshwellJ.D.LuF.W.VacchioM.S., 2000. Glucocorticoids in T cell development and function. Annu. Rev. Immunol.18:309.
45.
AgostiniM.Di MarcoB.NocentiniG.DelfinoD. V., 2002. Oxidative stress and apoptosis in immune diseases. Int. J. Immunopathol. Pharmacol.15:157.
46.
AcharyaA.2002. H2O2-induced apoptosis of thymocytes involves mobilization of divalent cations. Int. J. Immunopathol. Pharmacol.15:193.
47.
Di PietroR.CenturioneL.SabatiniN.BoscoD.SancilioS.GaraciF.RanaR.CataldiA., 2004. Caspase-3 is dually regulated by apoptogenic factors mitochondrial release and by SAPK/JNK metabolic pathway in leukemic cells exposed to etoposide-ionizing radiation combined treatment. Int. J. Immunopathol. Pharmacol.17:181.
48.
MarchisioM.GrimleyP.M.Di BaldassarreA.SantavenereE.MisciaS., 2004. Novel shift of Jak/Stat signaling characterizes the protective effects of aurintricarboxylic acid (ATA) from tumor necrosis factor-toxicity in human B lymphocytes. Int. J. Immunopathol. Pharmacol.17:5.
49.
JiangS.MeadowsJ.AndersonS.A.MukkadaA.J., 2002. Antileishmanial activity of the antiulcer agent omeprazole. Antimicrob. Agents Chemother. 46:2569.
50.
SuzukiK.TsuyuguchiK.MatsumotoH.NiimiA.TanakaE.AmitaniR., 2000. Effect of proton pump inhibitor alone or in combination with clarithromycin on mycobacterial growth in human alveolar macrophages. FEMS Microbiol. Lett.182:69.
51.
Zedtwitz-LiebensteinK.WenischC.PatrutaS.ParschalkB.DaxbockF.GraningerW., 2002. Omeprazole treatment diminishes intra- and extracellular neutrophil reactive oxygen production and bactericidal activity. Crit. Care Med. 30:1118.
52.
ScaringiL.CornacchioneP.FettucciariK.RosatiE.RossiR.MarconiP.CapodicasaE., 1996. Activity inhibition of cytolytic lymphocytes by omeprazole. Scand. J. Immunol.44:204.
53.
KataokaT.TakakuK.MagaeJ.ShinoharaN.TakayamaH.KondoS.NagaiK., 1994. Acidification is essential for maintaining the structure and function of lytic granules of CTL. Effect of concanamycin A, an inhibitor of vacuolar type H(+)-ATPase, on CTL-mediated cytotoxicity. J. Immunol.153:3938.
54.
YaoJ.ZhangG.J., 1996. Loss of lysosomal integrity caused by the decrease of proton translocation in methylene blue-mediated photosensitization. Biochim. Biophys. Acta.1284:35.
55.
ÖllingerK.2000. Inhibition of cathepsin D prevents free-radical-induced apoptosis in rat cardiomyocytes. Arch. Biochem. Biophys.373:346.
56.
ReinersJ.J.Jr.CarusoJ.A.MathieuP.ChelladuraiB.YinX.M.KesselD., 2002. Release of cytochrome c and activation of pro-caspase-9 following lysosomal photodamage involves Bid cleavage. Cell. Death Differ. 9:934.
57.
StokaV.TurkB.SchendelS.L.KimT.H.CirmanT.SnipasS.J.EllerbyL.M.BredesenD.FreezeH.AbrahamsonM.BrömineD.KrajewskiS.ReedJ.C.YinX.M.TurkV.SalvesenG.S., 2001. Lysosomal protease pathways to apoptosis. Cleavage of Bid, not pro-caspases, is the most likely route. J. Biol. Chem.276:3149.
58.
ZhaoM.EatonJ.W.BrunkU.T., 2000. Protection against oxidant-mediated lysosomal rupture: A new anti-apoptotic activity of Bcl-2?FEBS Lett. 485:104.
59.
RobergK.JohanssonU.ÖllingerK., 1999. Lysosomal release of cathepsin D precedes relocation of cytochrome c and loss of mitochondrial transmembrane potential during apoptosis induced by oxidative stress. Free Radic. Biol. Med.27:1228.
60.
BrunkU.T.NeuzilJ.EatonJ.W., 2001. Lysosomal involvement in apoptosis. Redox Rep. 6:91.
61.
SchotteP.DeclercqW.Van HuffelS.VandenabeeleP.BeyaertR., 1999. Non-specific effects of methyl ketone peptide inhibitors of caspases. FEBS Lett. 442:117.
62.
Di NapoliP.TaccardiA. A.GrilliA.FelacoM.Di GioacchinoL.De CaterinaR.BarsottiA., 2002. Verapamil reduces coronary endothelium damage and cardiomyocyte necrosis but not apoptosis after ischemia and reperfusion: Ex vivo study in rat hearts. Int. J. Immunopathol. Pharmacol.15:225.
63.
RobertsL.R.KurosawaH.BronS.F.FesmierP.J.AgellonL.B.LeungW.Y.MaoF.GoresG.J., 1997. Cathepsin B contributes to bile salt-induced apoptosis of rat hepatocytes. Gastroenterology113: 1714.
64.
VancompernolleK.Van HerrewegheF.PynaertG.Van de CraenM.De VosK.TottyN.SterlingA.FiersW.VandenabeeleP.GrootenJ., 1998. Atractyloside-induced release of cathepsin B, a protease with caspase-processing activity. FEBS Lett. 438:150.
65.
KrieserRJ.EastmanA., 1998. The cloning and expression of human deoxyribonuclease II. A possible role in apoptosis. J. Biol. Chem.273:30909.
