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Status |
Public on Apr 26, 2016 |
Title |
Gemcitabine Resistant Panc1 (Panc1-GR3) |
Sample type |
RNA |
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Source name |
Panc1 human pancreatic cancer cell line, gemcitabine resistant
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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
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Treatment protocol |
GEM-resistant cells were generated by exposure to gradually increasing concentrations of the reagent for 2 months. Parental Panc1 cells (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 Panc1 cells, and the three independent clones (Panc1-GRs: Panc1-GR1, -GR2, and –GR3) were used in the present experiments.
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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.
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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.
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Label |
Cy5
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Label protocol |
cDNA and Amino Allyl aRNA was synthesis by Amino Allyl MessageAmp II aRNA Amplification Kit (Ambion#1753). CyeDye Coupling and fragmentation were performed as the protocol supplied by TORAY Industries, Inc..
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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..
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Scan protocol |
3D-Gene Scanner ((Toray Industries Inc., Tokyo, Japan) was used for scanning. Images were quantified using Extraction(Toray Industries Inc., Tokyo, Japan).
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Description |
mRNA data expression from gemcitabine-resistant Panc1 cell clones
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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.
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Submission date |
Apr 25, 2016 |
Last update date |
Aug 22, 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 |
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Platform ID |
GPL13915 |
Series (1) |
GSE80617 |
mRNA Expression profiles of parental Panc cell lines and Gemcitabine resistance Panc1cell lines |
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