| Title |
Small nucleolar RNA U91 is a new internal control for accurate microRNAs quantification in pancreatic cancer
|
|---|---|
| Published in |
BMC Cancer, October 2015
|
| DOI | 10.1186/s12885-015-1785-9 |
| Pubmed ID | |
| Authors |
Alexey Popov, Arpad Szabo, Václav Mandys |
| Abstract |
RT-qPCR quantification of miRNAs expression may play an essential role in pancreatic ductal adenocarcinoma (PDAC) diagnostics. RT-qPCR-based experiments require endogenous controls for the result normalization and reliability. However, expression instability of reference genes in tumors may introduce bias when determining miRNA levels. We investigated expression of 6 miRNAs, isolated from FFPE samples of pancreatic adenocarcinomas. Four internal controls were utilized for RT-qPCR result normalization: artificial miR-39 from C. elegans, U6 snRNA, miR-16 and snoRNA U91. We found miR-21, miR-155 or miR-217 expression values in tumors may differ up to several times, depending on selected internal controls. Moreover, different internal controls can produce controversial results for miR-96, miR-148a or miR-196a quantification. Also, expression of our endogenous controls varied significantly in tumors. U6 demonstrated variation from -1.03 to 8.12-fold, miR-16 from -2.94 up to 7.38-fold and the U91 from -3.05 to 4.36-fold respectively. On the other hand, the most stable gene, determined by NormFinder algorithm, was U91. Each miRNA normalized relatively to the spike or U91, demonstrated similar expression values. Thus, statistically significant and insignificant differences between tumors and normal tissues for miRNAs were equal for the spike and the U91. Also, the differences between the spike and U91 were statistically insignificant for all of miRs except miR-217. Among three endogenous controls, U91 had the lowest average expression values and standard deviation in cancer tissues. We recommend U91 as a new normalizer for miRNA quantification in PDACs. |
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Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 1 | 100% |
Demographic breakdown
| Type | Count | As % |
|---|---|---|
| Members of the public | 1 | 100% |
Mendeley readers
The data shown below were compiled from readership statistics for 22 Mendeley readers of this research output. Click here to see the associated Mendeley record.
Geographical breakdown
| Country | Count | As % |
|---|---|---|
| Unknown | 22 | 100% |
Demographic breakdown
| Readers by professional status | Count | As % |
|---|---|---|
| Researcher | 5 | 23% |
| Student > Bachelor | 4 | 18% |
| Student > Ph. D. Student | 4 | 18% |
| Professor > Associate Professor | 3 | 14% |
| Student > Doctoral Student | 1 | 5% |
| Other | 4 | 18% |
| Unknown | 1 | 5% |
| Readers by discipline | Count | As % |
|---|---|---|
| Medicine and Dentistry | 8 | 36% |
| Agricultural and Biological Sciences | 4 | 18% |
| Biochemistry, Genetics and Molecular Biology | 2 | 9% |
| Pharmacology, Toxicology and Pharmaceutical Science | 1 | 5% |
| Immunology and Microbiology | 1 | 5% |
| Other | 1 | 5% |
| Unknown | 5 | 23% |
Attention Score in Context
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#20,294,248
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