title: A STAT3-decoy oligonucleotide induces cell death in a human colorectal carcinoma cell line by blocking nuclear transfer of STAT3 and STAT3-bound NF-κB.
abstract: BACKGROUND
  The transcription factor STAT3 (signal transducer and activator of transcription 3) is frequently activated in tumor cells. Activated STAT3 forms homodimers, or heterodimers with other TFs such as NF-κB, which becomes activated. Cytoplasmic STAT3 dimers are activated by tyrosine phosphorylation; they interact with importins via a nuclear localization signal (NLS) one of which is located within the DNA-binding domain formed by the dimer. In the nucleus, STAT3 regulates target gene expression by binding a consensus sequence within the promoter. STAT3-specific decoy oligonucleotides (STAT3-decoy ODN) that contain this consensus sequence inhibit the transcriptional activity of STAT3, leading to cell death; however, their mechanism of action is unclear.
  
  
  RESULTS
  The mechanism of action of a STAT3-decoy ODN was analyzed in the colon carcinoma cell line SW 480. These cells' dependence on activated STAT3 was verified by showing that cell death is induced by STAT3-specific siRNAs or Stattic. STAT3-decoy ODN was shown to bind activated STAT3 within the cytoplasm, and to prevent its translocation to the nucleus, as well as that of STAT3-associated NF-κB, but it did not prevent the nuclear transfer of STAT3 with mutations in its DNA-binding domain. The complex formed by STAT3 and the STAT3-decoy ODN did not associate with importin, while STAT3 alone was found to co-immunoprecipitate with importin. Leptomycin B and vanadate both trap STAT3 in the nucleus. They were found here to oppose the cytoplasmic trapping of STAT3 by the STAT3-decoy ODN. Control decoys consisting of either a mutated STAT3-decoy ODN or a NF-κB-specific decoy ODN had no effect on STAT3 nuclear translocation. Finally, blockage of STAT3 nuclear transfer correlated with the induction of SW 480 cell death.
  
  
  CONCLUSIONS
  The inhibition of STAT3 by a STAT3-decoy ODN, leading to cell death, involves the entrapment of activated STAT3 dimers in the cytoplasm. A mechanism is suggested whereby this entrapment is due to STAT3-decoy ODN's inhibition of active STAT3/importin interaction. These observations point to the high potential of STAT3-decoy ODN as a reagent and to STAT3 nucleo-cytoplasmic shuttling in tumor cells as a potential target for effective anti-cancer compounds.
authors.affiliation: INSERM, Unité 978, Bobigny, France.
authors.forename: Inès
authors.initials: I
authors.lastname: Souissi
authors.forename: Imen
authors.initials: I
authors.lastname: Najjar
authors.forename: Laurent
authors.initials: L
authors.lastname: Ah-Koon
authors.forename: Pierre Olivier
authors.initials: PO
authors.lastname: Schischmanoff
authors.forename: Denis
authors.initials: D
authors.lastname: Lesage
authors.forename: Stéphanie
authors.initials: S
authors.lastname: Le Coquil
authors.forename: Claudine
authors.initials: C
authors.lastname: Roger
authors.forename: Isabelle
authors.initials: I
authors.lastname: Dusanter-Fourt
authors.forename: Nadine
authors.initials: N
authors.lastname: Varin-Blank
authors.forename: An
authors.initials: A
authors.lastname: Cao
authors.forename: Valeri
authors.initials: V
authors.lastname: Metelev
authors.forename: Fanny
authors.initials: F
authors.lastname: Baran-Marszak
authors.forename: Remi
authors.initials: R
authors.lastname: Fagard
mesh_terms: D021581:Active Transport, Cell Nucleus; D016923:Cell Death; D045744:Cell Line, Tumor; D002466:Cell Nucleolus / Q000235:genetics / Q000378:metabolism*; D015179:Colorectal Neoplasms / Q000235:genetics / Q000378:metabolism* / Q000503:physiopathology*; D015972:Gene Expression Regulation, Neoplastic; D020868:Gene Silencing; D006801:Humans; D016328:NF-kappa B / Q000235:genetics / Q000378:metabolism*; D009841:Oligonucleotides / Q000235:genetics*; D011485:Protein Binding; D050796:STAT3 Transcription Factor / Q000235:genetics* / Q000378:metabolism*
chemical_list: D016328:NF-kappa B; D009841:Oligonucleotides; D050796:STAT3 Transcription Factor