Tumor Biology

, Volume 37, Issue 10, pp 14259–14269 | Cite as

Association of Sp1 and survivin in epithelial ovarian cancer: Sp1 inhibitor and cisplatin, a novel combination for inhibiting epithelial ovarian cancer cell proliferation

  • Umesh T. Sankpal
  • Susan B. Ingersoll
  • Sarfraz Ahmad
  • Robert W. Holloway
  • Vadiraja B. Bhat
  • Jerry W. Simecka
  • Liz Daniel
  • Ekamber Kariali
  • Jamboor K. Vishwanatha
  • Riyaz Basha
Original Article


The expression of specificity protein 1 (Sp1) and survivin was evaluated in clinical specimens of epithelial ovarian cancer (EOC) patients. When compared to normal tissue, EOC samples showed high expression of Sp1 and survivin using qPCR (Sp1: ∼2-fold; survivin: ∼5-fold) and Western blot (Sp1: >2.6-fold; survivin: >100-fold). The Sp1 inhibitor, and anti-cancer small molecule, tolfenamic acid (TA), was tested to enhance the response of Cisplatin (Cis) in EOC cell lines. Cell viability (CellTiter-Glo), combination index (CalcuSyn software), apoptosis (Annexin-V staining), cell cycle analyses (flow cytometry), and reactive oxygen species (flow cytometry) were determined. Cell migration and invasion was assessed using matrigel coated transwell chambers. Agilent Technologies proteomics analysis identified potential signaling pathways involved. The combination of TA (50 μM) and Cis (5 μM) synergistically increased the growth inhibition in ES2 (∼80 %, p < 0.001) and OVCAR-3 (60 %, p < 0.001) cells. TA or TA + Cis treatment in ES2 cells caused cell cycle arrest in G1 Phase (TA) or S-Phase (TA + Cis) and unregulated reactive oxygen species. Invasion and migration was decreased in ES2 cells. Global proteomic profiling showed modulation of proteins associated with oxidative phosphorylation, apoptosis, electron transport chain, DNA damage, and cell cycle proteins. These results demonstrate an association of Sp1 and survivin in EOC and confirm targeting these candidates with TA potentially sensitizes EOC cells to cisplatin.


Epithelial ovarian cancer Sp1 Survivin Cisplatin Tolfenamic acid 



This work was partially supported by a grant from the Ovarian Cancer Alliance of Florida (awarded to RB), Institute for Cancer Research and Pre-clinical Services, UNTHSC (RB & JWS), Florida Hospital Gala Endowed Program for Oncologic Research (SBI, SA & RWH). JKV is supported by a grant (1P20 MD006882) from the National Institute on Minority Health and Health Disparities, NIH.

Compliance with ethical standards


The information in this manuscript is based on either the published work or the opinion of the authors. The statements made in this manuscript not necessarily reflect the views of authors’ institutions.

Conflicts of interest



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Copyright information

© International Society of Oncology and BioMarkers (ISOBM) 2016

Authors and Affiliations

  • Umesh T. Sankpal
    • 1
  • Susan B. Ingersoll
    • 2
  • Sarfraz Ahmad
    • 2
  • Robert W. Holloway
    • 2
  • Vadiraja B. Bhat
    • 3
  • Jerry W. Simecka
    • 4
    • 5
  • Liz Daniel
    • 6
  • Ekamber Kariali
    • 7
  • Jamboor K. Vishwanatha
    • 1
    • 8
  • Riyaz Basha
    • 1
    • 4
    • 8
  1. 1.Institute for Cancer ResearchUniversity of North Texas Health Science CenterFort WorthUSA
  2. 2.Department of Gynecologic OncologyFlorida Hospital Cancer InstituteOrlandoUSA
  3. 3.Agilent Technologies, Inc.WilmingtonUSA
  4. 4.Pre-clinical ServicesUniversity of North Texas Health Science CenterFort WorthUSA
  5. 5.Pharmaceutical SciencesUniversity of North Texas Health Science CenterFort WorthUSA
  6. 6.MD Anderson Cancer Center OrlandoOrlandoUSA
  7. 7.Department of BiotechnologySambalpur UniversitySambalpurIndia
  8. 8.Molecular and Medical GeneticsUniversity of North Texas Health Science CenterFort WorthUSA

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