Combination of 13 cis-retinoic acid and tolfenamic acid induces apoptosis and effectively inhibits high-risk neuroblastoma cell proliferation

Sagar Shelake, Don Eslin, Robert M. Sutphin, Umesh T. Sankpal, Anmol Wadwani, Laura E. Kenyon, Leslie Tabor-Simecka, W. Paul Bowman, Jamboor K. Vishwanatha, Riyaz Basha

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Abstract

Chemotherapeutic regimens used for the treatment of Neuroblastoma (NB) cause long-term side effects in pediatric patients. NB arises in immature sympathetic nerve cells and primarily affects infants and children. A high rate of relapse in high-risk neuroblastoma (HRNB) necessitates the development of alternative strategies for effective treatment. This study investigated the efficacy of a small molecule, tolfenamic acid (TA), for enhancing the anti-proliferative effect of 13 cis-retinoic acid (RA) in HRNB cell lines. LA1-55n and SH-SY5Y cells were treated with TA (30. μM) or RA (20. μM) or both (optimized doses, derived from dose curves) for 48. h and tested the effect on cell viability, apoptosis and selected molecular markers (Sp1, survivin, AKT and ERK1/2). Cell viability and caspase activity were measured using the CellTiter-Glo and Caspase-Glo kits. The apoptotic cell population was determined by flow cytometry with Annexin-V staining. The expression of Sp1, survivin, AKT, ERK1/2 and c-PARP was evaluated by Western blots. The combination therapy of TA and RA resulted in significant inhibition of cell viability (. p<. 0.0001) when compared to individual agents. The anti-proliferative effect is accompanied by a decrease in Sp1 and survivin expression and an increase in apoptotic markers, Annexin-V positive cells, caspase 3/7 activity and c-PARP levels. Notably, TA. +. RA combination also caused down regulation of AKT and ERK1/2 suggesting a distinct impact on survival and proliferation pathways via signaling cascades. This study demonstrates that the TA mediated inhibition of Sp1 in combination with RA provides a novel therapeutic strategy for the effective treatment of HRNB in children.

Original languageEnglish
Pages (from-to)92-99
Number of pages8
JournalInternational Journal of Developmental Neuroscience
Volume46
DOIs
StatePublished - 1 Nov 2015

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Keywords

  • Chemotherapy
  • Infants
  • Neuroblastoma
  • Retinoic acid
  • Sympathetic nervous system
  • Tolfenamic acid

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