TY - JOUR
T1 - Combination of 13 cis-retinoic acid and tolfenamic acid induces apoptosis and effectively inhibits high-risk neuroblastoma cell proliferation
AU - Shelake, Sagar
AU - Eslin, Don
AU - Sutphin, Robert M.
AU - Sankpal, Umesh T.
AU - Wadwani, Anmol
AU - Kenyon, Laura E.
AU - Tabor-Simecka, Leslie
AU - Bowman, W. Paul
AU - Vishwanatha, Jamboor K.
AU - Basha, Riyaz
N1 - Funding Information:
This work is partially supported by the start-up funds to RB from the Institute for Cancer Research, UNT Health Science Center, Fort Worth, TX .
Publisher Copyright:
© 2015 Elsevier Ltd.
PY - 2015/11/1
Y1 - 2015/11/1
N2 - 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.
AB - 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.
KW - Chemotherapy
KW - Infants
KW - Neuroblastoma
KW - Retinoic acid
KW - Sympathetic nervous system
KW - Tolfenamic acid
UR - http://www.scopus.com/inward/record.url?scp=84940764954&partnerID=8YFLogxK
U2 - 10.1016/j.ijdevneu.2015.07.012
DO - 10.1016/j.ijdevneu.2015.07.012
M3 - Article
C2 - 26287661
AN - SCOPUS:84940764954
SN - 0736-5748
VL - 46
SP - 92
EP - 99
JO - International Journal of Developmental Neuroscience
JF - International Journal of Developmental Neuroscience
ER -