TY - JOUR
T1 - Calcium-regulatory proteins as modulators of chemotherapy in human neuroblastoma
AU - Florea, Ana Maria
AU - Varghese, Elizabeth
AU - McCallum, Jennifer E.
AU - Mahgoub, Safa
AU - Helmy, Irfan
AU - Varghese, Sharon
AU - Gopinath, Neha
AU - Sass, Steffen
AU - Theis, Fabian J.
AU - Reifenberger, Guido
AU - Büsselberg, Dietrich
N1 - Funding Information:
The authors would like to thank Safaa Shaheen and Noothan Jyothi Satheesh for their help and assistance with the FACS experiments for protein expression analysis. We thank the Flow Cytometry Facility within the Microscopy Core at Weill Cornell Medical College in Qatar for contributing to these studies. The Core is supported by the 'Biomedical Research Program at Weill Cornell Medical College in Qatar', a program funded by Qatar Foundation. We also would like to acknowledge the Genomics and Transcriptomics Core Facility at the Biological and Medical Research Center (BMFZ), Heinrich Heine University Düsseldorf, for performing the Affymetrix HuGene 2.0 ST chip hybridization and scanning.This study was supported by NPRP grant # 6-089-3-021 from the Qatar National Research Fund (a member of Qatar Foundation). The statements made herein are solely the responsibility of the authors.
PY - 2017
Y1 - 2017
N2 - Neuroblastoma (NB) is a pediatric cancer treated with poly-chemotherapy including platinum complexes (e.g. cisplatin (CDDP), carboplatin), DNA alkylating agents, and topoisomerase I inhibitors (e.g. topotecan (TOPO)). Despite aggressive treatment, NB may become resistant to chemotherapy. We investigated whether CDDP and TOPO treatment of NB cells interacts with the expression and function of proteins involved in regulating calcium signaling. Human neuroblastoma cell lines SH-SY5Y, IMR-32 and NLF were used to investigate the effects of CDDP and TOPO on cell viability, apoptosis, calcium homeostasis, and expression of selected proteins regulating intracellular calcium concentration ([Ca2+]i). In addition, the impact of pharmacological inhibition of [Ca2+]i-regulating proteins on neuroblastoma cell survival was studied. Treatment of neuroblastoma cells with increasing concentrations of CDDP (0.1-10 μM) or TOPO (0.1 nM-1 μM) induced cytotoxicity and increased apoptosis in a concentration- and time-dependent manner. Both drugs increased [Ca2+]i over time. Treatment with CDDP or TOPO also modified mRNA expression of selected genes encoding [Ca2+]i-regulating proteins. Differentially regulated genes included S100A6, ITPR1, ITPR3, RYR1 and RYR3. With FACS and confocal laser scanning microscopy experiments we validated their differential expression at the protein level. Importantly, treatment of neuroblastoma cells with pharmacological modulators of [Ca2+]i-regulating proteins in combination with CDDP or TOPO increased cytotoxicity. Thus, our results confirm an important role of calcium signaling in the response of neuroblastoma cells to chemotherapy and suggest [Ca2+]i modulation as a promising strategy for adjunctive treatment.
AB - Neuroblastoma (NB) is a pediatric cancer treated with poly-chemotherapy including platinum complexes (e.g. cisplatin (CDDP), carboplatin), DNA alkylating agents, and topoisomerase I inhibitors (e.g. topotecan (TOPO)). Despite aggressive treatment, NB may become resistant to chemotherapy. We investigated whether CDDP and TOPO treatment of NB cells interacts with the expression and function of proteins involved in regulating calcium signaling. Human neuroblastoma cell lines SH-SY5Y, IMR-32 and NLF were used to investigate the effects of CDDP and TOPO on cell viability, apoptosis, calcium homeostasis, and expression of selected proteins regulating intracellular calcium concentration ([Ca2+]i). In addition, the impact of pharmacological inhibition of [Ca2+]i-regulating proteins on neuroblastoma cell survival was studied. Treatment of neuroblastoma cells with increasing concentrations of CDDP (0.1-10 μM) or TOPO (0.1 nM-1 μM) induced cytotoxicity and increased apoptosis in a concentration- and time-dependent manner. Both drugs increased [Ca2+]i over time. Treatment with CDDP or TOPO also modified mRNA expression of selected genes encoding [Ca2+]i-regulating proteins. Differentially regulated genes included S100A6, ITPR1, ITPR3, RYR1 and RYR3. With FACS and confocal laser scanning microscopy experiments we validated their differential expression at the protein level. Importantly, treatment of neuroblastoma cells with pharmacological modulators of [Ca2+]i-regulating proteins in combination with CDDP or TOPO increased cytotoxicity. Thus, our results confirm an important role of calcium signaling in the response of neuroblastoma cells to chemotherapy and suggest [Ca2+]i modulation as a promising strategy for adjunctive treatment.
KW - Calcium signaling
KW - Chemotherapy
KW - Cisplatin
KW - Neuroblastoma
KW - Topotecan
UR - http://www.scopus.com/inward/record.url?scp=85016948961&partnerID=8YFLogxK
U2 - 10.18632/oncotarget.15283
DO - 10.18632/oncotarget.15283
M3 - Article
AN - SCOPUS:85016948961
SN - 1949-2553
VL - 8
SP - 22876
EP - 22893
JO - Oncotarget
JF - Oncotarget
IS - 14
ER -
