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Investigational New Drugs

, Volume 35, Issue 2, pp 158–165 | Cite as

Targeting specificity protein 1 transcription factor and survivin using tolfenamic acid for inhibiting Ewing sarcoma cell growth

  • Sagar Shelake
  • Umesh T. Sankpal
  • W. Paul Bowman
  • Matthew Wise
  • Anish RayEmail author
  • Riyaz BashaEmail author
PRECLINICAL STUDIES

Summary

Transcription factor Specificity protein 1 (Sp1) and its downstream target survivin (inhibitor of apoptosis protein), play major roles in the pathogenesis of various cancers. Ewing Sarcoma (ES) is a common soft tissue/bone tumor in adolescent and young adults. Overexpression of survivin is also linked to the aggressiveness and poor prognosis of ES. Small molecule Tolfenamic acid (TA) inhibits Sp1 and survivin in cancer cells. In this investigation, we demonstrate a strategy to target Sp1 and survivin using TA and positive control Mithramycin A (Mit) to inhibit ES cell growth. Knock down of Sp1 using small interfering RNA (siRNA) resulted in significant (p < 0.05) inhibition of CHLA-9 and TC-32 cell growth as assessed by CellTiter-Glo assay kit. TA or Mit treatment caused dose/time-dependent inhibition of cell viability, and this inhibition was correlated with a decrease in Sp1 and survivin protein levels in ES cells. Quantitative PCR results showed that Mit treatment decreased the mRNA expression of both survivin and Sp1, whereas TA diminished only survivin but not Sp1. Proteasome inhibitor restored TA-induced inhibition of Sp1 protein expression suggesting that TA might cause proteasome-dependent degradation. Gel shift assay using ES cell nuclear extract and biotinylated Sp1 consensus oligonucleotides confirmed that both TA and Mit decreased DNA-binding activity of Sp1. These results demonstrate that both Mit and TA reduce expression of Sp1 and survivin, disrupt Sp1 DNA-binding and inhibit ES cell proliferation. This investigation suggests that targeting Sp1 and survivin could be an effective strategy for inhibiting ES cell growth.

Keywords

Ewing sarcoma Sp1 Survivin Tolfenamic acid Mithramycin Targeted therapy 

Notes

Compliance with ethical standards

Conflict of interest

All authors (SS, UTS, WPW, MW, AR and RB) of this manuscript declare no conflict of interest.

Funding

This work is partially supported by the Institute for Cancer Research of UNT Health Science Center to RB and the Young Investigator research award from ‘Hyundai Hope on Wheels’ awarded to AR.

Ethical approval

Research presented in this manuscript does not contain studies with human participants or animals.

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Copyright information

© Springer Science+Business Media New York 2016

Authors and Affiliations

  1. 1.Department of PediatricsUniversity of North Texas Health Science CenterFort WorthUSA
  2. 2.Texas College of Osteopathic MedicineUniversity of North Texas Health Science CenterFort WorthUSA
  3. 3.Hematology and OncologyCook Children’s Medical CenterFort WorthUSA

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