|
| Status |
Public on Jul 01, 2011 |
| Title |
RKO aza PBA |
| Sample type |
RNA |
| |
|
| Source name |
RKO, 5-aza-C and 4-PBA
|
| Organism |
Homo sapiens |
| Characteristics |
disease state: colorectal cancer
cell line: RKO
treatment: 5-aza-2'-deoxycytidine plus 4-phenylbutyric acid
|
| Treatment protocol |
Cells were treated with 2 microM 5-aza-2’-deoxycytidine (aza) for 72 h, replacing the drug and medium every 24 h. In the combined protocol, cells were treated first with 2 microM aza for 72 h, and then with 3 mM 4-phenylbutyrate (PBA) for an additional 72 h, replacing the drug and medium every 24 h.
|
| Growth protocol |
HCT116 and DNMT double KO cells were cultured in McCoy 5A medium supplemented with 10% heat-inactivated fetal bovine serum. DLD1 and RKO cells were cultured in DMEM supplemented with 10% heat-inactivated fetal bovine serum. The cultures were incubated at 37 ºC in a humidified incubator with 5% CO2. Nomal colon RNA was purchased from BioChain.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was extracted using a mirVana miRNA isolation kit (Ambion).
|
| Label |
Cy3
|
| Label protocol |
Total RNA (100 ng) was labeled using a miRNA Labeling Reagent & Hybridization kit (Agilent Technologies) according to manufacturer's instruction.
|
| |
|
| Hybridization protocol |
Hybridization was performed using a miRNA Labeling Reagent & Hybridization kit (Agilent Technologies) according to manufacturer's instruction.
|
| Scan protocol |
Array was scanned with an Agilent G2565BA Microarray Scanner.
|
| Description |
miRNA expression in a human colorectal cancer cell line RKO treated with 5-aza-2'-deoxycytidine plus 4-phenylbutyric acid
|
| Data processing |
Agilent Feature Extraction Software (v 9.5) was used for background subtraction.
|
| |
|
| Submission date |
Jun 10, 2011 |
| Last update date |
Jul 01, 2011 |
| Contact name |
Hiromu Suzuki |
| E-mail(s) |
hsuzuki@sapmed.ac.jp
|
| Phone |
+81-11-611-2111
|
| Organization name |
Sapporo Medical University
|
| Department |
Department of Molecular Biology
|
| Street address |
S1, W17, Chuo-ku
|
| City |
Sapporo |
| State/province |
Hokkaido |
| ZIP/Postal code |
060-8556 |
| Country |
Japan |
| |
|
| Platform ID |
GPL6955 |
| Series (1) |
| GSE29900 |
microRNA expression signatures in colorectal cancer cell lines with demethylating treatment |
|
