|
| Status |
Public on Mar 09, 2016 |
| Title |
CMM1_Piroxicam_6h_rep2 |
| Sample type |
RNA |
| |
|
| Source name |
CMM1_Piroxicam_116 µM_6h
|
| Organism |
Canis lupus familiaris |
| Characteristics |
cell line: Mi/CMM1
cell type: canine melanoma cell line
cell line source: oral melanoma
treated with: piroxicam at 116 µMfor 6 h
|
| Treatment protocol |
CMM1 cell line was seeded at 6.3×10^3 cells/cm2. After 24 hours, piroxicam, carprofen and robenacoxib were added at their half maximal inhibitory concentration (779, 116 and 156 microM, respectively). Cells were harvested after another 24 hour-incubation.
|
| Growth protocol |
A canine melanoma cell line (CMM1) was maintained at 37°C in a humidified atmosphere of 5% CO2 in RPMI-1640 medium (Wako Pure Chemical Industries Ltd.,Osaka, Japan) supplemented with 10% heat-inactivated fetal bovine serum (FBS) (Gibco BRL, NY, USA) and 5 mg/L gentamicin (Sigma-Aldrich, MO, USA).
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Cells were lysed on dish and RNA toal RNA was ourified using RNeasy Mini Kit (Qiagen) according to the manufacturer’s instruction. RNA purity and integrity were confirmed with Agilent 2100 Bioanalyzer (Agilent Technologies). The RNA integrity numbers were above 9.0.
|
| Label |
Cy3
|
| Label protocol |
cDNA and Cy3-labeled cRNA were synthesized using Quick Amp Labeling Kit (Agilent Technologies) and RNA Spike-In One-Color Kit (Agilent Technologies) according to the manufacturer's instruction.
|
| |
|
| Hybridization protocol |
The purified cRNAs were quantified and hybridized to the Canine ver. 2 (4 × 44K) array (Agilent Technologies) using the Gene Expression Hybridization Kit (Agilent Technologies) according to the manufacturer's instruction.
|
| Scan protocol |
The hybridized array was washed with Gene Expression Wash (Agilent Technologies) and captured using a High-Resolution Microarray Scanner (Agilent Technologies) according to the manufacturer's instruction. Feature Extraction Software (Agilent Technologies) was used to analyze the scanned images.
|
| Description |
Piroxicam_2
Gene expression data of canine melanoma cell line after exposure to three different NSAIDs
|
| Data processing |
Probe annotation was performed using GeneSpring software (ver. 13.0) (Agilent technology). Data incorporation, preprocessing and quantile normalization were performed using R software (ver. 3.2.2) (R Development Core Team, 2005) and the limma R package (ver. 3.24.15)
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| |
|
| Submission date |
Mar 08, 2016 |
| Last update date |
Mar 09, 2016 |
| Contact name |
Kohei Saeki |
| E-mail(s) |
asaeki@mail.ecc.u-tokyo.ac.jp
|
| Phone |
+81358415405
|
| Organization name |
The University of Tokyo
|
| Street address |
1-1-1 Yayoi, Bunkyo
|
| City |
Tokyo |
| State/province |
Tokyo |
| ZIP/Postal code |
113-8657 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15379 |
| Series (1) |
| GSE78988 |
Change in gene expression of a canine melanoma cell line caused by exposure to nonsteroidal anti-inflammatory drugs (NSAIDs) . |
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