|
| Status |
Public on Sep 30, 2016 |
| Title |
ACHN |
| Sample type |
RNA |
| |
|
| Source name |
renal cancer cell lines_parental
|
| Organism |
Homo sapiens |
| Characteristics |
cell type: renal cancer cell lines
|
| Treatment protocol |
For RCC sunitinib resistant xenografts model, 6×10^6 cells of 786-O or ACHN cells were injected subcutaneously into the flanks of nude mice. When the volume of xenografts reached 200 mm3, mice were treated with sunitinib orally (40 mg/kg/day) in a standard 4 weeks on, 2 weeks off treatment schedule. After one cycle of treatment, RCC cells dissociated from xenografts were transplanted into secondary mice followed with the same sunitinib-treatment. Xenografts with sunitinib resistance were defined as unaffected xenograft growth under sunitinib pressure. Freshly isolated RCC cells obtained from the 3rd generation xenografts (termed 7Su3rd and ACSu3rd) were used for subsequent experiment.
|
| Growth protocol |
The RCC cell lines 786-O and ACHN (ATCC, USA), as well as 7Su3rd, ACSu3rd were cultured in DMEM (GIBCO, Invitrogen Inc, Carlsbad, CA, USA) supplemented with 10% fetal bovine serum (FBS, Gibco). 7Su3rd and ACSu3rd cells were chronically exposed in sunitinib at 5μM
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted from the specimens using Trizol reagent (Invitrogen) and purified with mirVana miRNA Isolation Kit (Ambion, Austin, TX, USA). RNA purity and concentration were determined from OD260/280 readings using spectrophotometer (NanoDrop ND-1000) and RNA integrity was determined by 1% formaldehyde denaturing gel electrophoresis.
|
| Label |
Cy3
|
| Label protocol |
cDNA labeled with a fluorescent dye (Cy3-dCTP) was produced by Eberwine’s linear RNA amplification method and subsequent enzymatic reaction.
|
| |
|
| Hybridization protocol |
The labeled RNA was purified and hybridized to microarrays according to the Agilent manufacturer's instructions.
|
| Scan protocol |
Images were scanned with the Agilent microarray scanner (Agilent 2565CA), gridded, and analyzed using Agilent feature extraction software version 10.10.
|
| Description |
Gene expression profiling of ACHN cell lines
|
| Data processing |
The scanned images were analyzed with Feature Extraction Software 10.10 (Agilent) using default parametersto obtain background subtracted and spatially detrended Processed Signal intensities. Features flagged in Feature Extraction as Feature Non-uniform outliers were excluded.
|
| |
|
| Submission date |
Jun 03, 2015 |
| Last update date |
Sep 30, 2016 |
| Contact name |
le qu |
| E-mail(s) |
[email protected]
|
| Organization name |
Second Military Medical University
|
| Department |
Department of Urology
|
| Street address |
440 North Chengdu Road
|
| City |
Shanghai |
| ZIP/Postal code |
200003 |
| Country |
China |
| |
|
| Platform ID |
GPL19748 |
| Series (1) |
| GSE69535 |
Gene expression profiling of generated sunitinib-resistant RCC cell lines |
|
