Abstract 1197
Background
Neuroblastoma (Nbl) is an embryonal tumour, originating from progenitor cells of the sympathetic nervous system, and is the most common extra-cranial solid tumour of childhood. Cisplatin is one of the most commonly used drugs in the treatment of Nbl. Unfortunately, the development of resistance against this cytotoxic agent limits its clinical use. We aimed to gain further understanding of the mechanisms underlying development of cisplatin resistance using an in vitro cell line model.
Methods
Cytotoxicity of cisplatin, carboplatin and oxaliplatin in neuroblastoma UKF-NB-4 and UKF-NB-4CDDP cells cultured in the exponential growth was determined. Peptide separations were carried out on an Easy-nLC 1000 nano system. MS analysis was performed using a Q-Exactive MS. MS data were analyzed with Proteome Discoverer (version1.4.1.14) using standardized workflows. The mass spectrum *.raw file was searched against the human SwissProt 57.15 database (20266 sequences protein entries) using MASCOT search engine (version 2.3, Matrix Science).
Results
The two cell lines that were used were the following: cisplatin-resistant UKF-NB4CDDP and UKF-NB4 cell lines. We proved decreased sensitivity of line UKF-NB-4CDDP to cisplatin and crossresitance to carboplatin and oxaliplatin. Out of a total 1802 identified proteins, 1448 were common expressed in two datasets. A total of 281 significant protein expressions were exclusively registered in the UKF-NB4CDDP cell line and 73 proteins were exclusively identified in UKF-NB4 cell line. UKF-NB-4CDDP and UKF-NB-4 cell lines showed overexpression of trafficking of transport in two datasets: comparison between of UKF-NB-4CDDP and UKF-NB-4 cell lines and proteins exclusive identified in UKF-NB-4CDDP. Exosome vesicles assay showed increase of vesicles in UKF-NB-4CDDP cell lines.
Conclusions
The Orbitap MS results could shed some light on the proteins involved in inducing resistance to cisplatin in cancer cells. These data strongly suggest that exosomes potently induce properties of Nbl cells and their chemoresistance to cisplatin and reinforces the potential benefit of trafficking of biological material across membranes in cancer.
Clinical trial identification
Legal entity responsible for the study
Mendel University in Brno, Czech Republic.
Funding
AZV project 15-28334A and Erasmus+: Staff Mobility - European Commission.
Editorial Acknowledgement
Disclosure
All authors have declared no conflicts of interest.
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