|
| Status |
Public on Oct 30, 2008 |
| Title |
ADH-1 ip saline ILI 48hrs post ILI |
| Sample type |
RNA |
| |
|
| Source name |
human-derived melanoma cells grown in rat as xenograft
|
| Organism |
Homo sapiens |
| Characteristics |
human melanoma-derived cell lines injected into hind limb of Crl:NIH Rnu Nude Rat
|
| Treatment protocol |
When tumors had reached ~1cm diameter treatment started. Animals were given either ADH-1 (100mg/kg) or saline via ip injection 1 hour prior to start of Isolated Limb Infusion (ILI) with either melphalan (LPAM) or saline. 4 hour time point samples were harvested 4 hours after the completion of ILI. All other animals were given a second dose of ADH-1 (100mg/kg) 24hrs after the first. 24 hour time point tissue was harvested 1 hour later. Remaining animals were given 2 more doses of ADH-1 (100mg/kg) over next 24hrs and tumor was harvested 1hr following final dose. Rats were anesthetized for tumor harvest and were humanely euthanized at the end of the study following an IACUC approved protocol. Harvested tumor was frozen immediately in liquid nitrogen and stored at -80C until ready for processing.
|
| Growth protocol |
Cells used to generate xenograft in rat were grown in Iscove MEM with 10%FBS, 1% Pen/Strep and 2mM L-glutamine added; 37C, 5% CO2; Cells were prepared in PBS plus matrigel at a ratio of 2:1 and 4 million cells were injected per rat.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
RNA was isolated using Qiagen Rneasy plus kit. RNA quality was assessed using Agilent bioanalyzer prior to labeling and hybridization.
|
| Label |
biotin
|
| Label protocol |
cDNA was synthesized from the total RNA and, using an Affymetrix recommended IVT protocol, biotin-labeled cRNA was generated from the cDNA. Prior to hybridization the cRNA was fragmented.
|
| |
|
| Hybridization protocol |
Cleaned and fragmented cRNA was hybridized to the Affymetrix Human U133 plus 2 genechip for 16 hrs at 45C followed by washing and staining according to Affymetrix instructions
|
| Scan protocol |
Hybridized probe arrays were stained according to Affymetrix protocols (streptavidin phycoerythrin conjugate) and scanned with the GeneChip Scanner
|
| Description |
gene expression data from melanoma xenografts following ILI with saline
|
| Data processing |
The amount of light emitted is directly proportional to the amount of target bound to each location on the probe array. Expression values were generated by calculating the difference between the perfect match signal and the mismatch signal for each probe using the Affymetrix GeneChip Operating System.
|
| |
|
| Submission date |
Jul 27, 2007 |
| Last update date |
Aug 28, 2018 |
| Contact name |
Christina Augustine |
| E-mail(s) |
christi.augustine@duke.edu
|
| Organization name |
Duke University Medical Center
|
| Department |
Surgery
|
| Street address |
VAMC RmE4001 508 Fulton St.
|
| City |
Durham |
| State/province |
NC |
| ZIP/Postal code |
27705 |
| Country |
USA |
| |
|
| Platform ID |
GPL570 |
| Series (1) |
| GSE8615 |
In vivo study of ADH-1 in combination with melphalan for treatment of melanoma |
|
| Relations |
| Reanalyzed by |
GSE119087 |
