Junction region of EWS-FLI1 fusion protein has a dominant negative effect in Ewing’s Sarcoma in vitro

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• 作者：Babu Jully (1)
  Ramshankar Vijayalakshmi (1)
  Gopisetty Gopal (1)
  Kesavan Sabitha (1)
  Thangarajan Rajkumar (1)
  
• 刊名：BMC Cancer
• 出版年：2012
• 出版时间：December 2012
• 年：2012
• 卷：12
• 期：1
• 全文大小：1134KB
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  35. The pre-publication history for this paper can be accessed here:http://www.biomedcentral.com/1471-2407/12/513/prepub
  
• 作者单位：Babu Jully (1)
  Ramshankar Vijayalakshmi (1)
  Gopisetty Gopal (1)
  Kesavan Sabitha (1)
  Thangarajan Rajkumar (1)
  
  1. Department of Molecular Oncology, Cancer Institute (WIA), 38, Sardar Patel Road, Chennai, 600036, India
  
• ISSN：1471-2407

文摘

Background Ewing’s sarcoma is a malignancy characterized by a specific 11:22 chromosomal translocation which generates a novel EWS-FLI1 fusion protein functioning as an aberrant transcription factor. In the present study, we have further characterized the junction region of the EWS-FLI1 fusion protein. Methods In-silico model of EWS-FLI1 fusion protein was analysed for ligand binding sites, and a putative region (amino acid (aa) 251-43 of the type 1 fusion protein) in the vicinity of the fusion junction was cloned and expressed using bacterial expression. The recombinant protein was characterized by Circular Dichroism (CD). We then expressed aa 251-80 ectopically in Ewing’s sarcoma cell-line and its effect on cell proliferation, tumorigenicity and expression of EWS-FLI1 target genes were analysed. Results Our modelling analysis indicated that Junction region (aa 251-43) encompasses potential ligand biding sites in the EWS-FLI1 protein and when expressed in bacteria was present as soluble form. Ectopically expressing this region in Ewing’s sarcoma cells inhibited tumorigenicity, and EWS-FLI1 target genes indicating a dominant negative biological effect. Conclusions Junction region can be exploited further as target for drug development in future to specifically target EWS-FLI1 in Ewing’s Sarcoma.