For investigation of the killing and proapoptotic effects of sodium salicylate (Na-Sal) on HL-60 cells, the cytotoxic activity of Na-Sal was measured by means of MTT assay. Apoptosis was identified and analyzed with the help of transmission electron microscopy, annexin V staining, and DNA gel electrophoresis, and the association of caspase-8 activation with apoptosis was determined with the specific protease inhibitor IETD-fmk. After exposure of HL-60 cells to increasing concentrations of NaSal (0.5,1,3,5, and 7 mmol/L) for 24 hours, the mean cell viability gradually dropped to 92%, 83%, 68%, 50%, and 42%. With treatment of target cells with 5-mmol/L (IC50) Na-Sal for 6,12, 24, or 36 hours, the mean cell survival tapered to 91%, 81%, 48%(P < .05 versus control), and 14%(P < .05 versus control). Again incubated with 5-mmol/L Na-Sal for 12 or 24 hours, HL-60 cells displayed clear early or late signs of apoptosis, including (1) notable enhancement of phosphatidylserine externalization, (2) cell shrinkage, membrane blebbing, and eventual disintegration into numerous apoptotic bodies, and (3) formation of ladder DNA. The viability of HL-60 cells increased significantly during 24 or 36 hours of coculture with 100-μmol/L IETD-fmk in combination with 5-mmol/L Na-Sal compared with the viability when 5-mmol/L Na-Sal was used alone(P < .05). Moreover, the target cells showed a considerable decrease in phosphatidylserine exposure and DNA fragmentation after coincubation for 12 or 24 hours performed as described above. The findings presented herein strongly suggest that Na-Sal can exert potent killing and proapoptotic activity against HL-60 cells, and this effect appears to depend on caspase-8 activation.
Sodium salicylate HL-60 cell line Apoptosis Kill Caspase-8
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Smalley WE, DuBois RN. Colorectal cancer and nonsteroidal anti-inflammatory drugs.Adv Pharmacol. 1997;39:1–20.CrossRefGoogle Scholar
Shiff SJ, Qiao L, Tsai LL, Rigas B. Sulindac sulfide, an aspirin-like compound, inhibits proliferation, causes cell cycle quiescence, and induces apoptosis in HT-29 colon adenocarcinoma cells.J Clin Invest. 1995;96:491–503.CrossRefPubMedPubMedCentralGoogle Scholar
Brunce CM, Mountford JC, French PJ, et al. Protection of myeloid differentiation by anti-inflammatory agents, by steroids and by retinoic acid involves a single intracellular target, probably an enzyme of the aldoketoreductase family.Biochem Biophys Acta. 1996;1311:189–198.CrossRefGoogle Scholar
Shiff SJ, Koutsos MI, Qiao L, Rigas B. Nonsteroidal anti-inflammatory drugs inhibit the proliferation of colon adenocarcinoma cells: effects on cell cycle and apoptosis.Exp Cell Res. 1996;222:179–188.CrossRefPubMedGoogle Scholar
Koopman G, Reulelingsperger CPM, Kuijten GAM, Keelmen RMJ, Pals ST, van Oers MHJ. Annexin V for flow cytometric detection of phosphatidylserine expression on B cells undergoing apoptoses.Blood. 1994;84:1415–1420.PubMedGoogle Scholar
van den Eijnde SM, Boshart L, Reutelingsperger CPM, De Zeeuw CI, Vermeijkeers C. Phosphatidylserine plasma membrane asymmetry in vivo: a pancellular phenomenon with alters during apoptosis.Cell Death Differ. 1997;4:311–316.CrossRefPubMedGoogle Scholar
van Engeland M, Ramaekers FCS, Schutte B, Reutelingsperger CPM. A novel assay to measure loss of plasma membrane asymmetry during apoptosis of adherent cells in culture.Cytometry. 1996;24:131–139.CrossRefPubMedGoogle Scholar
Slee EA, Harte MT, Kluch RM, et al. Ordering the cytochrome C— intitated caspase cascade: hierarchical activation of caspase-2,-3,-6,-7,-8, and -10 in a caspase-9-dependent manner.J Cell Biol. 1999;144:281–292.CrossRefPubMedPubMedCentralGoogle Scholar
Wickremasinghe RG, Hoffbrand AV. Biochemical and genetic con- trol of apoptosis: relevance to normal hematopoiesis and hematological malignancies [review].Blood. 1999;93:3587–3600.PubMedGoogle Scholar
Eischen CM, Kottke TJ, Martins LM, et al. Comparison of apoptosis in wild-type and Fas-resistant cells: chemotherapy-induced apoptosis is not dependent on Fas/Fas ligand interactions.Blood. 1997; 90:935–943.PubMedGoogle Scholar
Wesselborg S, Engels IH, Rossmann E, Los M, Schulze-Osthoff K. Anticancer drugs induce caspase-8/FLICE activation and apoptosis in the absence of CD95 receptor/ligand interaction.Blood. 1999;93:3053–3063.PubMedGoogle Scholar
Wieder T, Essmann F, Prokop A, et al. Activation of caspase-8 in drug-induced apoptosis of B-lymphoid cells is independent of CD95/Fas receptor-ligand interaction and occurs downstream of caspase-3.Blood. 2001;97:1379–1387.CrossRefGoogle Scholar