title: Knockdown of survivin (BIRC5) causes apoptosis in neuroblastoma via mitotic catastrophe.
abstract: BIRC5 (survivin) is one of the genes located on chromosome arm 17q in the region that is often gained in neuroblastoma. BIRC5 is a protein in the intrinsic apoptotic pathway that interacts with XIAP and DIABLO leading to caspase-3 and caspase-9 inactivation. BIRC5 is also involved in stabilizing the microtubule-kinetochore dynamics. Based on the Affymetrix mRNA expression data, we here show that BIRC5 expression is strongly upregulated in neuroblastoma compared with normal tissues, adult malignancies, and non-malignant fetal adrenal neuroblasts. The over-expression of BIRC5 correlates with an unfavorable prognosis independent of the presence of 17q gain. Silencing of BIRC5 in neuroblastoma cell lines by various antisense molecules resulted in massive apoptosis as measured by PARP cleavage and FACS analysis. As both the intrinsic apoptotic pathway and the chromosomal passenger complex can be therapeutically targeted, we investigated in which of them BIRC5 exerted its essential anti-apoptotic role. Immunofluorescence analysis of neuroblastoma cells after BIRC5 silencing showed formation of multinucleated cells indicating mitotic catastrophe, which leads to apoptosis via P53 and CASP2. We show that BIRC5 silencing indeed resulted in activation of P53 and we could rescue apoptosis by CASP2 inhibition. We conclude that BIRC5 stabilizes the microtubules in the chromosomal passenger complex in neuroblastoma and that the apoptotic response results from mitotic catastrophe, which makes BIRC5 an interesting target for therapy.
authors.affiliation: Department of Oncogenomics, Academic Medical Center, University of Amsterdam, Meibergdreef 15, PO Box 22700, 1105 AZ Amsterdam, The Netherlands.
authors.forename: Fieke
authors.initials: F
authors.lastname: Lamers
authors.forename: Ida
authors.initials: I
authors.lastname: van der Ploeg
authors.forename: Linda
authors.initials: L
authors.lastname: Schild
authors.forename: Marli E
authors.initials: ME
authors.lastname: Ebus
authors.forename: Jan
authors.initials: J
authors.lastname: Koster
authors.forename: Bo R
authors.initials: BR
authors.lastname: Hansen
authors.forename: Troels
authors.initials: T
authors.lastname: Koch
authors.forename: Rogier
authors.initials: R
authors.lastname: Versteeg
authors.forename: Huib N
authors.initials: HN
authors.lastname: Caron
authors.forename: Jan J
authors.initials: JJ
authors.lastname: Molenaar
mesh_terms: D017209:Apoptosis / Q000502:physiology*; D015153:Blotting, Western; D053143:Caspase 2 / Q000502:physiology; D061945:Caspase Inhibitors; D045744:Cell Line, Tumor; D002470:Cell Survival / Q000502:physiology; D003546:Cysteine Endopeptidases / Q000502:physiology; D005434:Flow Cytometry; D015972:Gene Expression Regulation, Neoplastic; D057809:HEK293 Cells; D006801:Humans; D051034:Inhibitor of Apoptosis Proteins / Q000172:deficiency* / Q000235:genetics / Q000378:metabolism; D008938:Mitosis / Q000502:physiology*; D009447:Neuroblastoma / Q000235:genetics / Q000378:metabolism / Q000473:pathology*; D020411:Oligonucleotide Array Sequence Analysis / Q000379:methods; D009841:Oligonucleotides / Q000494:pharmacology; D012334:RNA, Neoplasm / Q000737:chemistry / Q000235:genetics; D034741:RNA, Small Interfering / Q000008:administration & dosage / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D000077022:Survivin; D046888:Tissue Array Analysis / Q000379:methods; D016159:Tumor Suppressor Protein p53 / Q000502:physiology
chemical_list: C107489:BIRC5 protein, human; D061945:Caspase Inhibitors; C552302:EZN 3042; D051034:Inhibitor of Apoptosis Proteins; D009841:Oligonucleotides; D012334:RNA, Neoplasm; D034741:RNA, Small Interfering; D000077022:Survivin; C495901:TP53 protein, human; D016159:Tumor Suppressor Protein p53; C490360:CASP2 protein, human; D053143:Caspase 2; D003546:Cysteine Endopeptidases