Repression of BIRC5/survivin by FOXO3/FKHRL1 sensitizes human neuroblastoma cells to DNA damage-induced apoptosis.

The phosphatidylinositol 3-kinase (PI3K)-protein kinase B (PKB) pathway regulates survival and chemotherapy resistance of neuronal cells, and its deregulation in neuroblastoma (NB) tumors predicts an adverse clinical outcome. Here, we show that inhibition of PI3K-PKB signaling in human NB cells induces nuclear translocation of FOXO3/FKHRL1, represses the prosurvival protein BIRC5/Survivin, and sensitizes to DNA-damaging agents. To specifically address whether FKHRL1 contributes to Survivin regulation, we introduced a 4-hydroxy-tamoxifen-regulated FKHRL1(A3)ERtm allele into NB cells. Conditional FKHRL1 activation repressed Survivin transcription and protein expression. Transgenic Survivin exerted a significant antiapoptotic effect and prevented the accumulation of Bim and Bax at mitochondria, the loss of mitochondrial membrane potential as well as the release of cytochrome c during FKHRL1-induced apoptosis. In concordance, Survivin knockdown by retroviral short hairpin RNA technology accelerated FKHRL1-induced apoptosis. Low-dose activation of FKHRL1 sensitized to the DNA-damaging agents doxorubicin and etoposide, whereas the overexpression of Survivin diminished FKHRL1 sensitization to these drugs. These results suggest that repression of Survivin by FKHRL1 facilitates FKHRL1-induced apoptosis and sensitizes to cell death induced by DNA-damaging agents, which supports the central role of PI3K-PKB-FKHRL1 signaling in drug resistance of human NB.

[1]  D. Altieri,et al.  A novel anti-apoptosis gene, survivin, expressed in cancer and lymphoma , 1997, Nature Medicine.

[2]  G P Nolan,et al.  High-efficiency gene transfer and selection of human hematopoietic progenitor cells with a hybrid EBV/retroviral vector expressing the green fluorescence protein. , 1998, Cancer research.

[3]  D. Altieri,et al.  Anti-apoptosis gene, survivin, and prognosis of neuroblastoma , 1998, The Lancet.

[4]  M. Ohira,et al.  High expression of Survivin, mapped to 17q25, is significantly associated with poor prognostic factors and promotes cell survival in human neuroblastoma , 2000, Oncogene.

[5]  S. Agrawal,et al.  Survivin inhibition induces human neural tumor cell death through caspase‐independent and ‐dependent pathways , 2001, Journal of neurochemistry.

[6]  K. Ihara,et al.  Quick quantitative analysis of gene dosages associated with prognosis in neuroblastoma. , 2001, Cancer letters.

[7]  J. Lammers,et al.  FKHR-L1 can act as a critical effector of cell death induced by cytokine withdrawal , 2002, The Journal of cell biology.

[8]  Wenhua Zheng,et al.  FKHRL1 and its homologs are new targets of nerve growth factor Trk receptor signaling , 2002, Journal of neurochemistry.

[9]  P. Coffer,et al.  FOXO transcription factors directly activate bim gene expression and promote apoptosis in sympathetic neurons , 2003, The Journal of cell biology.

[10]  G Böck,et al.  Cyclin D3 and c-MYC control glucocorticoid-induced cell cycle arrest but not apoptosis in lymphoblastic leukemia cells , 2004, Cell Death and Differentiation.

[11]  D. Grossman,et al.  Rapid induction of mitochondrial events and caspase-independent apoptosis in Survivin-targeted melanoma cells , 2004, Oncogene.

[12]  John Calvin Reed,et al.  An IAP-IAP Complex Inhibits Apoptosis* , 2004, Journal of Biological Chemistry.

[13]  D. Chung,et al.  Phosphatidylinositol 3-kinase inhibition down-regulates survivin and facilitates TRAIL-mediated apoptosis in neuroblastomas. , 2004, Journal of pediatric surgery.

[14]  J. Jaboin,et al.  Genetic and pharmacologic identification of Akt as a mediator of brain-derived neurotrophic factor/TrkB rescue of neuroblastoma cells from chemotherapy-induced cell death. , 2005, Cancer research.

[15]  Rogier Versteeg,et al.  Biological effects of TrkA and TrkB receptor signaling in neuroblastoma. , 2005, Cancer letters.

[16]  T. Lörch,et al.  Automatic telomere length measurements in interphase nuclei by IQ‐FISH , 2005, Cytometry. Part A : the journal of the International Society for Analytical Cytology.

[17]  E. Beierle,et al.  VEGF-mediated survivin expression in neuroblastoma cells. , 2005, The Journal of surgical research.

[18]  W. London,et al.  Survivin mRNA Levels Are Associated With Biology of Disease and Patient Survival in Neuroblastoma: A Report From the Children's Oncology Group , 2006, Journal of pediatric hematology/oncology.

[19]  J. Meléndez-Zajgla,et al.  Regulation of mitochondrial Smac/DIABLO-selective release by survivin , 2007, Oncogene.

[20]  S. Fulda,et al.  Activation of Akt predicts poor outcome in neuroblastoma. , 2007, Cancer research.

[21]  D. Altieri,et al.  Compartmentalized phosphorylation of IAP by protein kinase A regulates cytoprotection. , 2007, Molecular cell.

[22]  C. Thiele,et al.  Downregulation of Bim by brain-derived neurotrophic factor activation of TrkB protects neuroblastoma cells from paclitaxel but not etoposide or cisplatin-induced cell death , 2007, Cell Death and Differentiation.

[23]  D. Tindall,et al.  Dynamic FoxO transcription factors , 2007, Journal of Cell Science.

[24]  B. Burgering,et al.  Stressing the role of FoxO proteins in lifespan and disease , 2007, Nature Reviews Molecular Cell Biology.

[25]  B. Meister,et al.  FKHRL1-mediated expression of Noxa and Bim induces apoptosis via the mitochondria in neuroblastoma cells , 2007, Cell Death and Differentiation.