Inhibition of NF-kappa B can enhance Fas-mediated apoptosis in leukemia cell line HL-60

Abstract

This study explored the effects of nuclear factor-kappa B (NF-κB) inhibitor Bay 11-7082 on Fas/FasL system and Fas-mediated apoptosis in cell line HL-60 cells. The mRNA and protein levels of Fas, FasL, and X-linked inhibitor of apoptosis protein (XIAP) were detected by reverse transcription-polymerase chain reaction (RT-PCR) and flow cytometry (FCM); the level of sFasL was evaluated by enzyme-linked immunosorbent assay (ELISA); and apoptosis was determined by FCM. After treatment with Bay 11-7082, the mRNA and protein levels of FasL and XIAP in HL-60 cells were significantly lower than in the controls (P<0.05), but the mRNA and protein levels of Fas and sFasL did not change significantly (P>0.05). Apoptotic rate of HL-60 cells treated with Bay 11-7082 was significantly higher than in the controls (P<0.05). Therefore, we conclude that Bay 11-7082 can enhance Fas-mediated apoptosis in HL-60 cells by downregulating FasL and XIAP levels.

Keywords

nuclear factor-kappa B / Fas/FasL system / HL-60 / Bay 11-7082

References

[1] FriesenC, HerrI, KrammerP H, DebatinK M. Involvement of the CD95(APO-1/Fas)receptor/ligand system in drug-induced apoptosis in leukaemia cell. Nature Med, 1996, 2(5): 574-577

[2] KimK M, LeeK, HongY S, ParkH Y. Fas-mediated apoptosis and expression of related genes in human malignant hematopoietic cell. Exp Mol Med, 2000, 32(4): 246-254

[3] KordesU, KrappmannD, HeissmeyerV, LudwigW D, ScheidereitC. Transcription factor NF-κB is constitutively activated in acute lymphoblastic leukemia cells. Leukemia, 2000, 14(3): 399-402

[4] QinY, Camoretti-MercadoB, BlokhL, LongC G, KoF D, HamannK J. Fas Resistance of Leukemic Eosinophils Is Due to Activation of NF-κB by Fas Ligation. J Immunol, 2002, 169(7): 3536-3544

[5] Bueso-RamosC E, RochaF C, ShishodiaS, MedeirosL J, KantarjianH M, Vadhan-RajS, EstrovZ, SmithT L, NguyenM H, AggarwalB B. Expression of constitutively active nuclear-kappa B RelA transcription factor in blasts of acute myeloid leukemia. Hum Pathol, 2004, 35(2): 246-253

[6] HafezM, Al-TonbaryY, El-BayoumiM A, HatemN, HawasS, MansourA, MarzoukI, HafezM M, YahiaS, FarahatN. Markers of apoptosis and proliferation related gene products as predictors of treatment outcome in childhood acute lymphoblastic leukemia. Hematology, 2007, 12(3): 209-218

[7] QinY, Camoretti-MercadoB, BlokhL, LongC G, KoF D, HamannK J. Fas Resistance of Leukemic Eosinophils Is Due to Activation of NF-κB by Fas Ligation. J Immunol, 2002, 169(7): 3536-3544

[8] BarnhartB C, PietrasE M, Algeciras-SchimnichA, SalmenaL, SayamaK, HakemR, PeterM E. CD95 apoptosis resistance in certain cells can be overcome by noncanonical activation of caspase-8. Cell Death Differ, 2005, 12(1): 25-37

[9] MeliM, D'AlessandroN, TolomeoM, RausaL, NotarbartoloM, DusonchetL. NF-κB Inhibiton Restores Sensitivity to Fas-Mediated Apoptosis in Lymphoma Cell Lines. Ann N Y Acad Sci, 2003, 1010: 232-236

[10] BuzynA, PetitF, OstankovitchM, FigueiredoS, VaretB, GuilletJ G, AmeisenJ C, EstaquierJ. Membrane-bound Fas ligand on leukemic cells:a mechanism of tumor immune escape in leukemic patients. Blood, 1999, 94(9): 3135-3140

[11] BarnhartB C, LegembreP, PietrasE, BubiciC, FranzosoG, PeterM E. CD95 ligand induces motility and invasiveness of apoptosis-resistant tumor cells. EMBO J, 2004, 23(15): 3175-3185

[12] KavurmaMM, KhachigianLM. Signaling and transcriptional control of Fas ligand gene expression. Cell Death Differ, 2003, 10(1): 36-44

[13] SavickieneJ, TreigyteG, MagnussonK E, NavakauskieneR. p21 and FasL gene activation via Sp1 and NF-κB is required for leukemia cell survival but not for cell death induced by diverse stimuli. Int J Biochem Cell Biol, 2005, 37(4): 784-796

[14] KaterA P, DickerF, MangiolaM, WelshK, HoughtenR, OstreshJ, NefziA, ReedJ C, PinillaC, KippsT J. Inhibitors of XIAP sensitize CD40-activated chronic lymphocytic leukemia cells to CD95-mediated apoptosis. Blood, 2005, 106(5): 1742-1748

[15] LoederS, DrensekA, JeremiasI, DebatinK M, FuldaS. Small molecule XIAP inhibitors sensitize childhood acute leukemia cells for CD95-induced apoptosis. Int J Cancer, 2009, 126(9): 2216-2228

[16] DoyleB T, O'NeillA J, NewsholmeP, FitzpatrickJ M, WatsonR W. The loss of IAP expression during HL-60 cell differentiation is caspase-independent. J Leukoc Biol, 2002, 71(2): 247-254

[17] OhashiM, IwaseM, NagumoM. Changes in susceptibility to Fas-mediated apoptosis during differentiation of HL-60 cells. J Leukoc Biol, 2000, 67(3): 374-380

[18] HaoZ, MakT W. Type I and Type II pathways of Fas-mediated apoptosis are differentially controlled by XIAP. J Mol Cell Biol, 2009<month>Oct</month><day>16</day> [Epub ahead of print]

[19] Straszewski-ChavezS L, AbrahamsV M, AldoP B, RomeroR, MorG. AKT controls human first trimester trophoblast cell sensitivity to FAS-mediated apoptosis by regulating XIAP expression. Biol Reprod, 2010, 82(1): 146-152

[20] BlombergJ, RuuthK, SantosD, LundgrenE. Acquired resistance to Fas/CD95 ligation in U937 cells is associated with multiple molecular mechanisms. Anticancer Res, 2008, 28(2A): 593-599

[21] QinY, AuhS, BlokhL, LongC, GagnonI, HamannK J. TNF-alpha induces transient resistance to Fas-induced apoptosis in eosinophilic acute myeloid leukemia cells. Cell Mol Immunol, 2007, 4(1): 43-52

[22] FaklerM, LoederS, VoglerM, SchneiderK, JeremiasI, DebatinK M, FuldaS. Small molecule XIAP inhibitors cooperate with TRAIL to induce apoptosis in childhood acute leukemia cells and overcome Bcl-2-mediated resistance. Blood, 2009, 113(8): 1710-1722

[23] MoralesJ C, Ruiz-MaganaM J, Ruiz-RuizC. Regulation of the resistance to TRAIL-induced apoptosis in human primary T lymphocytes: role of NF-kappaB inhibition. Mol Immunol, 2007, 44(10): 2587-2597

[24] CarterB Z, MakD H, SchoberW D, DietrichM F, PinillaC, VassilevL T, ReedJ C, AndreeffM. Triptolide sensitizes AML cells to TRAIL-induced apoptosis via decrease of XIAP and p53-mediated increase of DR5. Blood, 2008, 111(7): 3742-3750

Acknowledgements

We thank Jian-hua CHEN, Research Institute of Hematology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, for the provision of experimental facilities and technical support in the research work.

RIGHTS & PERMISSIONS

2014 Higher Education Press and Springer-Verlag Berlin Heidelberg