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| Status |
Public on Nov 03, 2015 |
| Title |
MicroRNA screening in human pancreatic carcinoma cell lines |
| Platform organism |
synthetic construct |
| Sample organism |
Homo sapiens |
| Experiment type |
Non-coding RNA profiling by array
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| Summary |
No reliable predictors of susceptibility to gemcitabine chemotherapy exist in pancreatic ductal adenocarcinoma. MicroRNAs (miR) are epigenetic gene regulators with tumorsuppressive or oncogenic roles in various carcinomas. This study assesses chemoresistant PDAC for its specific miR expression pattern.
Gemcitabine-resistant variants of two mutant p53 human pancreatic adenocarcinoma cell lines were established. MicroRNA screening was investigated by microarray.
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| Overall design |
Gemcitabine-resistant PANC-1 (PANC-1-GR) and MIA-PaCa-2 (MIA-PaCa-2-GR) cell clones were produced by exposing the parental cells to repeated pulsatile gemcitabine treatment over 3 days with constant sublethal concentrations followed by recovery-periods with agent-free medium until the cells recovered exponentially. Parental PANC-1 cells were treated with 0.4µM gemcitabine cycles for approximately 9 months. Parental MIA-PaCa-2 cells were exposed to 0.06µM gemcitabine cycles for approximately 12 months. Affymetrix GeneChip miRNA microarrays (Affymetrix UK Ltd., High Wycombe, UK) were performed in parental and chemoresistant PANC-1 and MIA-PaCa-2 cells after 29 chemotherapy cycles using the manufacturers´ protocols. The samples were prepared from 1µg of total-RNA in accordance with the Affymetrix FlashTag Biotin HSR RNA Labeling Kit. The targets were hybridized overnight to Affymetrix GeneChip miRNA arrays. Following hybridization, the arrays were washed and stained using the Affymetrix GeneChip Fluidics Station 450 and scanned using the Affymetrix GeneChip Scanner 3000 7G. Microarray data quality was checked as recommended by the manufacturer and by the quality metrics in the Partek Genomics Suite software (Partek Inc., St. Louis, MO).
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| Contributor(s) |
Dhayat SA, Seggewiß J |
| Citation(s) |
26606261 |
| Submission date |
Nov 02, 2015 |
| Last update date |
Jul 29, 2019 |
| Contact name |
Sameer Dhayat |
| E-mail(s) |
sameer.dhayat@ukmuenster.de
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| Phone |
00492518351402
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| Organization name |
Comprehensive Cancer Center Muenster
|
| Street address |
Albert-Schweitzer-Campus 1 ( W1)
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| City |
Muenster |
| ZIP/Postal code |
48149 |
| Country |
Germany |
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| Platforms (1) |
| GPL16384 |
[miRNA-3] Affymetrix Multispecies miRNA-3 Array |
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| Samples (12)
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| GSM1923061 |
Gemcitabine-resistant Mia-PaCa-2_biological p29_1 |
| GSM1923062 |
Gemcitabine-resistant Mia-PaCa-2_biological p29_2 |
| GSM1923063 |
Gemcitabine-resistant Mia-PaCa-2_biological p29_3 |
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| This SubSeries is part of SuperSeries: |
| GSE74574 |
microRNA screening in human pancreatic adenocarcinoma |
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| Relations |
| BioProject |
PRJNA300841 |
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