|
| Status |
Public on Dec 01, 2017 |
| Title |
ISL-treated (Cy5)/Control (Cy3)-PC-3 |
| Sample type |
RNA |
| |
|
| Channel 1 |
| Source name |
ISL-treated PC-3 cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: PC-3 Prostate Cancer Cell Line
treatment: ISL
|
| Treatment protocol |
LNCaP cells and PC-3 cells were seeded into 25 cm2 flask at a density of 5×104 cells per flask. After 24h of incubation, cells were treated with 25 μM ISL or 0.025% DMSO as control for 48h.
|
| Growth protocol |
LNCaP cells or PC-3 cells were maintained in RPMI-1640 (Hyclone, USA) supplemented with 10% FBS (Hyclone, USA), 1% penicillin and streptomycin at 37°C in a humidified incubator containing 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After treatment, cells were harvested and washed with cold PBS. Total RNA was extracted using Trizol Reagent (Invitrogen, USA) according to the manufacturer's instrctions and stored at -80°C waiting for quality test and microarray analysis. The quality test and microarray experiment were performed by CapitlBio Corporation (Beijing, China). Total RNA was purified using NucleoSpin RNA clean-up kit (MACHEREY-NAGEL, Germany) and quantified by a spectrophotometer. The quality of samples were tested through electrophoresis on gel containing formaldehyde. Samples passing the qualifty test were allowed to step into the next stage of the experiment.
|
| Label |
Cy5
|
| Label protocol |
The labeling process was performed by using Jingxin cRNA Amplification and Labeling Kit according to the manufacture's instructions.
|
| |
|
| Channel 2 |
| Source name |
Control PC-3 cells
|
| Organism |
Homo sapiens |
| Characteristics |
cell line: PC-3 Prostate Cancer Cell Line
treatment: Control (untreated)
|
| Treatment protocol |
LNCaP cells and PC-3 cells were seeded into 25 cm2 flask at a density of 5×104 cells per flask. After 24h of incubation, cells were treated with 25 μM ISL or 0.025% DMSO as control for 48h.
|
| Growth protocol |
LNCaP cells or PC-3 cells were maintained in RPMI-1640 (Hyclone, USA) supplemented with 10% FBS (Hyclone, USA), 1% penicillin and streptomycin at 37°C in a humidified incubator containing 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
After treatment, cells were harvested and washed with cold PBS. Total RNA was extracted using Trizol Reagent (Invitrogen, USA) according to the manufacturer's instrctions and stored at -80°C waiting for quality test and microarray analysis. The quality test and microarray experiment were performed by CapitlBio Corporation (Beijing, China). Total RNA was purified using NucleoSpin RNA clean-up kit (MACHEREY-NAGEL, Germany) and quantified by a spectrophotometer. The quality of samples were tested through electrophoresis on gel containing formaldehyde. Samples passing the qualifty test were allowed to step into the next stage of the experiment.
|
| Label |
Cy3
|
| Label protocol |
The labeling process was performed by using Jingxin cRNA Amplification and Labeling Kit according to the manufacture's instructions.
|
| |
|
| |
| Hybridization protocol |
Labeled DNA was hybridized in 2X GEx Hyb Buffer (HI-RPM) containing 25% formamide at 45°C overnight. After hybridization, microarrays were washed sequentially.
|
| Scan protocol |
Microarrays were scanned on an Agilent G2565CA Microarray Scanner.
|
| Description |
single sample, using qRT-PCR to validate the microarray result
|
| Data processing |
Agilent Feature Extraction Software was used for image quantification, background subtraction and LOWESS mormalization.
|
| |
|
| Submission date |
Jul 19, 2017 |
| Last update date |
Jan 23, 2018 |
| Contact name |
Biyan Zhang |
| E-mail(s) |
zhang_bi_yan@hotmail.com
|
| Organization name |
Shanghai University of Traditional Chinese Medicine
|
| Department |
School of Basic Medicine
|
| Street address |
1200 Cai Lun Road, Pudong District
|
| City |
Shanghai |
| State/province |
Shanghai |
| ZIP/Postal code |
201203 |
| Country |
China |
| |
|
| Platform ID |
GPL17077 |
| Series (1) |
| GSE101640 |
Human Prostate Cancer Cells: ISL-treated cells vs. Control |
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