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

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Tumor Biology

Abstract

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.

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Acknowledgments

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.

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Authors and Affiliations

  1. Institute for Cancer Research, University of North Texas Health Science Center, Fort Worth, TX, 76107, USA

    Umesh T. Sankpal, Jamboor K. Vishwanatha & Riyaz Basha

  2. Department of Gynecologic Oncology, Florida Hospital Cancer Institute, Orlando, FL, 32804, USA

    Susan B. Ingersoll, Sarfraz Ahmad & Robert W. Holloway

  3. Agilent Technologies, Inc., Wilmington, DE, 19808, USA

    Vadiraja B. Bhat

  4. Pre-clinical Services, University of North Texas Health Science Center, Fort Worth, TX, 76107, USA

    Jerry W. Simecka & Riyaz Basha

  5. Pharmaceutical Sciences, University of North Texas Health Science Center, Fort Worth, TX, 76107, USA

    Jerry W. Simecka

  6. MD Anderson Cancer Center Orlando, Orlando, FL, 32806, USA

    Liz Daniel

  7. Department of Biotechnology, Sambalpur University, Jyoti Vihar, Sambalpur, Odisha, 768019, India

    Ekamber Kariali

  8. Molecular and Medical Genetics, University of North Texas Health Science Center, Fort Worth, TX, 76107, USA

    Jamboor K. Vishwanatha & Riyaz Basha

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  1. Umesh T. Sankpal
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Correspondence to Riyaz Basha.

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Sankpal, U.T., Ingersoll, S.B., Ahmad, S. et al. Association of Sp1 and survivin in epithelial ovarian cancer: Sp1 inhibitor and cisplatin, a novel combination for inhibiting epithelial ovarian cancer cell proliferation. Tumor Biol. 37, 14259–14269 (2016). https://doi.org/10.1007/s13277-016-5290-9

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  • DOI: https://doi.org/10.1007/s13277-016-5290-9

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