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Journal: Science translational medicine
Article Title: Interfering with Resistance to Smoothened Antagonists by Inhibition of the PI3K Pathway in Medulloblastoma
doi: 10.1126/scitranslmed.3001599
Figure Lengend Snippet: RNA from sensitive tumors treated with either vehicle or NVP-LDE225 (20mg/kg/day) for 4, 16 and 48 hours, and resistant tumors treated with 20 or 160 mg/kg/day (mpk) for 26 days (d) was profiled on Affymetrix mouse gene expression arrays. A, Pattern of genes initially inhibited by NVP-LDE225 but reemerging in resistant tumors. Numbers express relative rank in expression. B, Top ranking genes by Spearman correlation of expression matching with the pattern in 2A. C, Heat map of the top ranked genes associated with the pattern in 2A. Each gene’s expression values are z-transformed for comparability, with red indicating relatively high expression and green indicating relatively low expression. D, CGH analysis using the Agilent Mouse Genome CGH Microarray Kit 244A of three resistant (LDE 1,2,3) and three sensitive (Veh 1,2,3) tumors identified a focal amplification in the region containing Gli2 on Chromosome 1 in 2 out of 3 resistant tumors. Copy number changes are expressed as log2. E, Additional DNA from normal liver (NL), vehicle (controls) and resistant tumors was analyzed by quantitative PCR for the Gli2 locus. F, Correlation between levels of Gli2 mRNA expression and Gli2 copy number is shown for vehicle-treated control tumors (open square) and resistant tumors that emerged after treatment with NVP-LDE225 at 10 bid (green square), 20 qd (red diamond), 40 bid (brown triangle), 80 bid (blue circle) and 160 bid (black inverse triangle) mg/kg/day. Axis in log10 scale. G, Inhibition of Gli2 mRNA levels by siRNA knock-down of Gli2 was associated with decreased proliferation and Gli1 mRNA expression in resistant Gli2 amplified (copy number:20) and Gli2-overexpressing (17-fold) medulloblastoma tumors. Two independent Gli2 siRNAs (Gli2-1 and Gli2-1) were used.
Article Snippet: D , CGH analysis using the
Techniques: Expressing, Transformation Assay, Microarray, Amplification, Real-time Polymerase Chain Reaction, Inhibition