The effect of rosi̇glatazone on di̇fferanti̇aton of neural stem cells in minimal residual disease model of neuroblastoma

Neuroblastoma (NB) is an embryonic tumor originating from the neural crest. The most important problem in advanced stage disease with clinical remission is the presence of minimal residual disease (MRD).Peroxisome proliferator-activating receptor is a nuclear transcription factor that regulates cell proliferation and differentiation.Rosiglatazone (RZG) peroxisome proliferator-activating receptor agonist is an antidiabetic agent. The aim of this study was to establish the MRD model and the determine whether rosiglatazone has any effect on the differantiation of neuronal stem cell in minimal residual disease model. In this study N-Myc positive KELLY and N-Myc negative SH-SY5Y cell lines were used. By analysing the viability of the cells WST-1 and IC50 doses were determined.NB cells were generated by MRH model based on residual cells with repeated cisplatin (CDDP) tretmeants.A 2-fold or greater increase/decrease in PHOX2B gene expression was used as an indicator that the cells reached the minimal residual disease model.PHOX2B gene expression was determined by RT-PCR method.The effects of cisplatin (CDDP) and RZG cell viability, apoptosis and neuronal stem cell differentiation of neuroblastoma cells in the MRD model were investigated by using S-100, Oct-3, Nestin, Sox2 and CD133 protein expression.The statistical analysis was performed using the non-parametric Mann-Whitney-U test in the SPSS22 program. p <0.05 was considered statistically significant. In SH-SY5Y and KELLY cells RZG IC50 doses were determined as 50,100µM; CDDP IC90 doses were determined as 80µM in KELLY and 50µM in SH-SY5Y. 400µM CDDP and CDDP+RGZ combinations reduced cell viability in NB cell lines and induced cells to apoptosis.In the cell cycle, most of the cells were imprisoned during the SubG1 phase. CD133+ in resistant cells.not observed.Resistive MRD KELLY and SH-SY5Y cell according to the control; in CDDP + RGZ combinations, expression of PHOX2B gene and OCT3-4 increased and Sox2 expression decreased. The combination of CDDP+RZG in SH-SY5Y cells with MRD model showed more apoptotic effect when compared with CDDP, decrease in differentiation markers, observed opposite situation in KELLY cells, and effect of RZG in relation to neuronal differentiation. Increased CD133+ cell counts in resistant MRH SH-SY5Y cells compared to non-resistant Showed resistance to CDDP and CD133+ cancer stem cells in MRD model.The emergence of different CD133 cell ratios in N-MYC (-) and (+) NB cells supported the fact that neuroblastoma was a heterogeneous tumor and this heterogeneity was encountered with stem cell profile. Rosiglatazone should be evaluate as an agent with CDDP main therapautic of neuroblastoma and as an supportive agent at PPRgama expresing tumors in MRD in-vivo NB model.