Abstract
The p73 gene codes for various different protein isoforms. They include proteins expressed under the control of the P1 promoter that contain a transactivation domain and are similar in function to p53 (TAp73 isoforms), as well as proteins regulated by the P2 promoter that lack this domain and function as dominant negative inhibitors of TAp73 and p53 (ΔNp73 isoforms). Whereas TAp73 functions as a tumor suppressor with pro-apoptotic function, ΔNp73 is likely to prevent the induction of apoptosis in tumor cells and to participate in oncogenesis. Here we used a loss-of-function strategy to assess the role of ΔNp73 in SH-SY5Y neuroblastoma cells. An antisense oligonucleotide designed to target ΔNp73 mRNA, but not TAp73, was used to effectively downregulate this transcript. ΔNp73 downregulation was accompanied by increased levels of the pro-apoptotic BH3 family member PUMA at the mRNA and protein level, and by conformational activation of BAX which translocated to mitochondria. These ΔNp73 antisense-mediated alterations led to the induction of apoptosis as detected by decreased cell viability, augmented DNA fragmentation and increased caspase-3 activity in cell lysates. Our results demonstrate the cytoprotective role of ΔNp73 in neuroblastoma and suggest its use as a target for molecular intervention therapy.
Similar content being viewed by others
References
KH Vousden X Lu (2002) ArticleTitleLive or let die: the cell’s response to p53 Nat Rev Cancer 2 594–604 Occurrence Handle10.1038/nrc864 Occurrence Handle1:CAS:528:DC%2BD38XlslGqsL8%3D Occurrence Handle12154352
A Yang N Walker R Bronson M Kaghad M Oosterwegel J Bonnin C Vagner H Bonnet P Dikkes A Sharpe F McKeon D Caput (2000) ArticleTitlep73-deficient mice have neurological, pheromonal and inflammatory defects but lack spontaneous tumours Nature 404 99–103
G Melino LV De KH Vousden (2002) ArticleTitlep73: Friend or foe in tumorigenesis Nat Rev Cancer 2 605–615
G Melino F Bernassola M Ranalli K Yee WX Zong M Corazzari RA Knight DR Green C Thompson KH Vousden (2003) ArticleTitlep73 induces apoptosis via PUMA transactivation and Bax mitochondrial translocation J Biol Chem 279 8076–8083
D Goldschneider E Blanc G Raguenez M Barrois A Legrand G Le Roux H Haddada J Benard S Douc-Rasy (2004) ArticleTitleDifferential response of p53 target genes to p73 overexpression in SH-SY5Y neuroblastoma cell line J Cell Sci 117 293–301
T Nakagawa M Takahashi T Ozaki K Watanabe S Hayashi M Hosoda S Todo A Nakagawara (2003) ArticleTitleNegative autoregulation of p73 and p53 by ΔNp73 in regulating differentiation and survival of human neuroblastoma cells Cancer Lett 197 105–109
TJ Grob U Novak C Maisse D Barcaroli AU Luthi F Pirnia B Hugli HU Graber LV De MF Fey G Melino A Tobler (2001) ArticleTitleHuman ΔNp73 regulates a dominant negative feedback loop for TAp73 and p53 Cell Death Differ 8 1213–1223
O Ishimoto C Kawahara K Enjo M Obinata T Nukiwa S Ikawa (2002) ArticleTitlePossible oncogenic potential of ΔNp73: a newly identified isoform of human p73 Cancer Res 62 636–641
T Stiewe S Zimmermann A Frilling H Esche BM Putzer (2002) ArticleTitleTransactivation-deficient ΔTA-p73 acts as an oncogene Cancer Res 62 3598–3602
AI Zaika N Slade SH Erster C Sansome TW Joseph M Pearl E Chalas UM Moll (2002) ArticleTitleΔNp73, a dominant-negative inhibitor of wild-type p53 and TAp73, is up-regulated in human tumors J Exp Med 196 765–780
S Vossio E Palescandolo N Pediconi F Moretti C Balsano M Levrero A Costanzo (2002) ArticleTitleDN-p73 is activated after DNA damage in a p53-dependent manner to regulate p53-induced cell cycle arrest Oncogene 21 3796–3803
S Allart H Martin C Detraves J Terrasson D Caput C Davrinche (2002) ArticleTitleHuman cytomegalovirus induces drug resistance and alteration of programmed cell death by accumulation of ΔN-p73α J Biol Chem 277 29063–29068
T Nakagawa M Takahashi T Ozaki K Watanabe Ki S Todo H Mizuguchi T Hayakawa A Nakagawara (2002) ArticleTitleAutoinhibitory regulation of p73 by ΔNp73 to modulate cell survival and death through a p73-specific target element within the ΔNp73 promoter Mol Cell Biol 22 2575–2585
MM Noesel Particlevan R Versteeg (2004) ArticleTitlePediatric neuroblastomas: genetic and epigenetic ‘Danse Macabre’ Gene 325 1–15
I Casciano K Mazzocco L Boni G Pagnan B Banelli G Allemanni M Ponzoni GP Tonini M Romani (2002) ArticleTitleExpression of ΔNp73 is a molecular marker for adverse outcome in neuroblastoma patients Cell Death Differ 9 246–251
A Ziegler AP Simões-Wüst U Zangemeister-Wittke (2000) ArticleTitleOptimizing efficacy of antisense oligodeoxynucleotides targeting inhibitors of apoptosis Meth Enzymol 314 477–490
AP Simões-Wüst RA Olie O Gautschi SH Leech R Haner J Hall D Fabbro RA Stahel U Zangemeister-Wittke (2000) ArticleTitleBcl-xl antisense treatment induces apoptosis in breast carcinoma cells Int J Cancer 87 582–590
J Gong F Traganos Z Darzynkiewicz (1994) ArticleTitleA selective procedure for DNA extraction from apoptotic cells applicable for gel electrophoresis and flow cytometry Anal Biochem 218 314–319
Z Darzynkiewicz G Juan X Li W Gorczyca T Murakami F Traganos (1997) ArticleTitleCytometry in cell necrobiology: analysis of apoptosis and accidental cell death (necrosis) Cytometry 27 1–20
O Matzura A Wennborg (1996) ArticleTitleRNAdraw: an integrated program for RNA secondary structure calculation and analysis under 32-bit Microsoft Windows Comput Appl Biosci 12 247–249
MT McManus PA Sharp (2002) ArticleTitleGene silencing in mammals by small interfering RNAs Nat Rev Genet 3 737–747
S Cory DC Huang JM Adams (2003) ArticleTitleThe Bcl-2 family: roles in cell survival and oncogenesis Oncogene 22 8590–8607 Occurrence Handle10.1038/sj.onc.1207102 Occurrence Handle1:CAS:528:DC%2BD3sXptFOmtb0%3D Occurrence Handle14634621
N Dias CA Stein (2002) ArticleTitleAntisense oligonucleotides: basic concepts and mechanisms Mol Cancer Ther 1 347–355
DA Braasch DR Corey (2002) ArticleTitleNovel antisense and peptide nucleic acid strategies for controlling gene expression Biochemistry 41 4503–4510
AA Levin (1999) ArticleTitleA review of the issues in the pharmacokinetics and toxicology of phosphorothioate antisense oligonucleotides Biochim Biophys Acta 1489 69–84
M Mihara S Erster A Zaika O Petrenko T Chittenden P Pancoska UM Moll (2003) ArticleTitlep53 has a direct apoptogenic role at the mitochondria Mol Cell 11 577–590 Occurrence Handle10.1016/S1097-2765(03)00050-9 Occurrence Handle1:CAS:528:DC%2BD3sXjtVWktrk%3D Occurrence Handle12667443
H Puthalakath A Strasser (2002) ArticleTitleKeeping killers on a tight leash: transcriptional and post-translational control of the pro-apoptotic activity of BH3-only proteins Cell Death Differ 9 505–512
Author information
Authors and Affiliations
Corresponding author
Rights and permissions
About this article
Cite this article
Simões-Wüst, A., Sigrist, B., Belyanskaya, L. et al. ΔNp73 antisense activates PUMA and induces apoptosis in neuroblastoma cells. J Neurooncol 72, 29–34 (2005). https://doi.org/10.1007/s11060-004-3118-8
Issue Date:
DOI: https://doi.org/10.1007/s11060-004-3118-8