|
| Status |
Public on Apr 26, 2016 |
| Title |
Gemcitabine Resistant Panc1 (Panc1-GR1) |
| Sample type |
RNA |
| |
|
| Source name |
Panc1 human pancreatic cancer cell line, gemcitabine resistant
|
| Organism |
Homo sapiens |
| Characteristics |
treatment: Gemcitabine resistant Panc1 derivative generated from the parental Panc1 line by consecutive treatments with 30ng/ml gemcitabine over a period of 2 months
|
| Treatment protocol |
GEM-resistant cells were generated by exposure to gradually increasing concentrations of the reagent for 2 months. Parental Panc1cells (Panc1-P) were exposed to GEM at an initial concentration of 2 ng/ml. When the cells adapted, the GEM concentration was increased. The final GEM concentration was 30 ng/ml. Using this process, we successfully generated GEM-resistant cells. Limiting the dilution of the established cells allowed the cloning of the GEM-resistant Panc1cells, and the three independent clones (Panc1-GRs: Panc1-GR1, -GR2, and –GR3) were used in the present experiments.
|
| Growth protocol |
Cells were cultured in Dulbecco’s modified Eagle’s medium supplemented with 10% fetal bovine serum (FBS) with antibiotics and incubated at 37 C in a humidified incubator with 5% CO2.
|
| Extracted molecule |
total RNA |
| Extraction protocol |
Total RNA was isolated from parental Panc1 and gemcitabine resistance Panc1 cell lines using Trizol reagent (Invitrogen) according to the manufacturer’s instructions.
|
| Label |
Hy5
|
| Label protocol |
Extracted total RNA was labeled with Hy5 using the miRCURY LNA Array miR labeling kit (Exiqon, Vedbaek, Denmark)
|
| |
|
| Hybridization protocol |
Hybridized for 16 h at 37 C with rotary shake (250 rpm). Hybridization buffer and washing protocol was followed by the protocol supplied by TORAY Industries, Inc..
|
| Scan protocol |
3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
|
| Description |
MicroRNA data expression from gemcitabine-resistant Panc1 cell clones
|
| Data processing |
The raw data of each spot was normalized by substitution with a mean intensity of the background signal determined by all blank spots’ signal intensities of 95% confidence intervals. Measurements of both duplicate spots with the signal intensities greater than 2 standard deviations (SD) of the background signal intensity were considered to be valid. A relative expression level of a given miRNA was calculated by comparing the signal intensities of the averaged valid spots with their mean value throughout the microarray experiments. 3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
|
| |
|
| Submission date |
Apr 25, 2016 |
| Last update date |
Apr 26, 2016 |
| Contact name |
Satoshi Kondo |
| Organization name |
Toray Industries,Inc.
|
| Department |
New Projects Development Division
|
| Street address |
Tebiro 6-10-1
|
| City |
Kamakura |
| State/province |
Kanagawa |
| ZIP/Postal code |
248-8555 |
| Country |
Japan |
| |
|
| Platform ID |
GPL15446 |
| Series (1) |
| GSE80616 |
microRNA Expression profiles of parental Panc1 cell lines and Gemcitabine resistance Panc1 cell lines |
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