Expression data from SQ-diEG treated mice

Global gene expression profiling reveals a potential anti-melanoma effect of SQ-diEG in B16F10 lung colonization mice model. Squalene (SQ) was considered as a promising natural agent in anti-cancer treatment due to its strong antioxidant and anti-inflammatory activity; however, its pharmacological value has been largely underestimated because of its poor solubility and bioavailability. To adress this problem, a novel amphiphilic SQ derivative which bearing ethylene glycol oligomers was synthesized and was used as a permeation enhancer in this study to check its potential effect on anti-melanoma metastasis using B16F10 lung colonization mice model. Total 40 six years old male C57BL/6J mice were randomly divided into five groups (8 mice per group), including the control "(-)B16F10 injected and water treated"; the model "(+) B16F10 injected and water treated"; the positive control "(+) B16F10 injected and DTIC treated (Dacarbazine, 70 mg/kg/day)"; the SQ-diEG treatment "(+) B16F10 injected and SQ-diEG treated (5 mg/kg/day)"; the SQ-diEG treatment "(+) B16F10 injected and SQ-diEG treated (25 mg/kg/day)". After one week of acclimatization, B16F10 were murine melanoma cells line which intravenously injected (200000 tumor cells in 100 μl PBS, 0.2 ml/mouse) through the tail vein of mice. Mice was orally administrated with Dacarbazine (DTIC, positive control, 70mg/kg), and SQ-diEG (5mg/kg and 25mg/kg) for 20 days. On the last day of administration, mice was sacrificed and its lung was washed and collect for the further RNA extraction. Finally, the mice microarray gene expression profiling was conducted for two biological replicates. RNA from the lung tissue was extracted using Isogen (Nippon Gene Co. Ltd., Toyama, Japan). The integrity of RNA was quantified using NanoDrop 2000 spectrophotometer (Thermo Scientific, Wilmington, DE, USA). RNA samples were prepared for gene expression profiling analysis with GeneChip® 3' Expression Arrays using 3' IVT PLUS Reagent Kit (Affymetrix Inc., Santa Clara, CA, USA). Two hundred and fifty ng of total RNA from each sample was used to generate amplified and biotinylated complementary RNA (cRNA) from poly (A) RNA in a total RNA sample according to the user manual. IVT Incubation time was 16 hour. GeneAtlas® Hybridization, Wash and Stain Kit was used for hybridizing 3' IVT Array Strips according to the user manual (P/N 08-0306). The mouse genome array strips were hybridized for 16 hours in a 45oC incubator, washed and stained and finally imaging was done with the GeneAtlas Fluidics and Imaging Station.