Gene expression array of neuroblastoma human tumors in NSG upon treatment with CXCR4 inhibitor BL8040

Purpose: CXCR4 plays a role in neuroblastoma development. The goal of this study is to search for pathways controlled by CXCR4 which contribute to neuroblastoma tumor development. Methods: Human SK-N-BE(2) cells were orthotopically injected into the left adrenal gland of 6-8 weeks old mice. During treatment protocol mice were monitored for 21 days. As of day 7, mice were treated with a daily injection of the CXCR4 inhibitor BL-8040 (400µg) for 14 consecutive days. Tumor RNA was generated from 3 mice in each group using Trizol. Amplified and biotinylated sense-strand DNA were prepared according to the standard Affymetrix protocol from 200ng total RNA (Expression Analysis Technical Manual, 2005, Affymetrix). Following fragmentation, 5.5µg of Biotinylated ssDNA were hybridized for 17 hr at 45ºC on GeneChip Human Gene 1.0 ST Array. GeneChip were washed and stained in the Affymetrix Fluidic Station 450. GeneChips were scanned using Affymetrix GeneChip Scanner 3000. Results: 98 genes were differentially expressed in the treated tumors. Conclusions: Inhibition of CXCR4 prevented tumor growth and reduced survival of tumor cells by significantly altering several apoptotic and cell cycle signaling pathways. NSG mice injected with SK-N-BE(2) cells to the adrenal gland. One week following cell injection mice were treated with BL-8040 for 14 consecutive days. Tumors were harvested and RNA purified from 3 control and treated mice were analyzed using the GeneChip Human Gene 1.0 ST Array and scanned with the Affymetrix GeneChip scanner 3000.