Transcriptomic Landscape of Cisplatin-Resistant Neuroblastoma Cells

Abstract

The efficiency of cisplatin (CDDP) is significantly hindered by the development of resistance during the treatment course. To gain a detailed understanding of the molecular mechanisms underlying the development of cisplatin resistance, we comparatively analyzed established a CDDP-resistant neuroblastoma cell line (UKF-NB-4CDDP) and its susceptible parental cells (UKF-NB-4). We verified increased chemoresistance of UKF-NB-4CDDP cells by analyzing the viability, induction of apoptosis and clonal efficiency. To shed more light on this phenomenon, we employed custom cDNA microarray (containing 2234 probes) to perform parallel transcriptomic profiling of RNA and identified that 139 genes were significantly up-regulated due to CDDP chemoresistance. The analyses of molecular pathways indicated that the top up-regulation scoring functions were response to stress, abiotic stimulus, regulation of metabolic process, apoptotic processes, regulation of cell proliferation, DNA repair or regulation of catalytic activity, which was also evidenced by analysis of molecular functions revealing up-regulation of genes encoding several proteins with a wide-spectrum of enzymatic activities. Functional analysis using lysosomotropic agents chloroquine and bafilomycin A1 validated their potential to re-sensitize UKF-NB-4CDDP cells to CDDP. Taken together, the identification of alterations in specific genes and pathways that contribute to CDDP chemoresistance may potentially lead to a renewed interest in the development of novel rational therapeutics and prognostic biomarkers for the management of CDDP-resistant neuroblastoma.

Metallothionein-3 promotes cisplatin chemoresistance remodelling in neuroblastoma

“…In addition, the up-regulation of ATP-binding cassette transporters (ABC family members) provides a supporting mechanism for direct efflux of CDDP from the cell via energy-fuelled active transport. CDDP can further activate the proteasome complex pathway, which was also demonstrated by our previous results 24 , 46 . Interestingly, we also identified a pronounced up-regulation of numerous proteasome subunits in UKF-NB-4 cells overexpressing hMT3.…”

“…CDDP is a cytotoxic drug that kills cancer cells by damaging DNA and inhibiting its synthesis 22 , 23 . Despite the high efficiency of CDDP, cancer cells frequently develop a CDDP-chemoresistance phenotype, which could be due to a wide spectrum of causes, including decreased blood flow into tumour mass, inhibited internalization of CDDP, exocytosis of CDDP, intracellular sequestration of CDDP, halted DNA repair or apoptosis-driving mechanisms, or a presence of quiescent and non-cycling cells (non-growing or non-dividing state for a long period of time) 24 . As shown in our previous transcriptomic study, up-regulation of hMT3 induced development of CDDP-chemoresistance in Nbl cells and significantly altered the expression of sets of genes involved in apoptosis and oncogene-induced senescence 25 .…”

Amino Acids Regulate Cisplatin Insensitivity in Neuroblastoma

“…Thus, utilizing cell lines as models mechanisms mediated through intra-tumoral oncogenes, mutations and signaling cascades, as well as extra-tumoral factors, such as stromal components and non-cellular components, have been identified to affect CDDP sensitivity [ 10 , 14 , 15 , 16 , 17 ]. Multiple mechanisms mediated by oncogenes, such as exosomal-microRNAs and cell signaling pathways, were identified to promote CDDP-insensitivity in neuroblastoma [ 10 , 18 , 19 , 20 ]. The oncogenes, including v-myc avian myelocytomatosis viral oncogene neuroblastoma derived homolog (MYCN) and Hypoxia-inducible factor (HIF)-1∝, were shown to regulate metabolism and mediate CDDP-resistance in neuroblastoma [ 21 , 22 , 23 ].…”

Drug Sequestration in Lysosomes as One of the Mechanisms of Chemoresistance of Cancer Cells and the Possibilities of Its Inhibition

“…In support of a role for MT-3 in lysosomal function, the absence of MT-3 results in changes in the levels of lysosomal proteins and results in reduced lysosomal degradative capacity. Lee et al, proposed that MTs might similarly stabilize lysosomes following autophagocytotic delivery to the lysosomal compartment [ 86 ]. Also, Cho et al demonstrated the roles of MT-3 and zinc in radiation-induced autophagy and radio-resistance in glioma cells [ 87 ].…”