Effect of ILB on Gene expression in SHSY5Y neuroblastoma cells. Effect of ILB on Gene expression in SHSY5Y neuroblastoma cells

The experiments were carried out to further understand the mechanism of action of ILB, a low molecular weight dextran sulphate, in neuronal cell. The experiments were carried out with ILB concentrations that are relevant to the therapeutic concentrations of the drug. The results indicate that ILB regulates the expression of genes to enhance cell survival and differentiation is a dose dependent manner. Upstream regulator analysis indicates that the gene expression changes are due to enhanced activity of growth factors and cytokines. Overall design: Human SHSY5Y neuroblastoma cells (CRL-2266, ATCC) were cultured in DMEM:F12 medium supplemented with 10% FCS and 1% GlutaMAX™-I for 48 hours (day 0 timepoint). ILB was added to the cultures (final concentration: 0.01, 0.1, 1.0 and 10 µg/ml; n=3 for each concentration). Control cultures received only vehicle (0.9% NaCl in water added to culture medium 1:10,000; n=2). Cells were cultured for another 48 hours (day 2 timepoint). Samples were preserved in RNAlater. RNA was extracted using the Agilent RNA Isolation Kit. Expression analysis was performed using the SurePrint G3 Human Unrestricted GE 8x60K array (G4858A- 028004).