Transcriptomics,Multiomics

Dataset Information

14

Reference Genes for MiRNAome Analysis of Urothelial Carcinomas


ABSTRACT: Background/Objective: Quantitative real-time PCR (RT-qPCR) is widely used in miRNA expression studies on cancer. To compensate for the analytical variability produced by the multiple steps of the method, relative quantification of the measured miRNAs is required, which is based on normalization to endogenous reference genes. A literature search in PubMed revealed that no study has been performed so far on reference miRNAs for normalization of miRNA expression in urothelial carcinoma. The aim of this study was to identify suitable reference genes for miRNA expression studies by RT-qPCR in urothelial carcinoma. Methods: Candidate reference miRNAs were selected from 24 papillary urothelial carcinoma and normal bladder tissue samples by miRNA microarrays. The usefulness of these candidate reference miRNAs together with the commonly for normalization purposes used small nuclear RNAs RNU6B, RNU48, and Z30 were thereafter validated by RT-qPCR in 58 tissue samples and analyzed by the algorithms geNorm, NormFinder, and BestKeeper. Principal Findings: Based on the miRNA microarray data, a total of 16 miRNAs were identified as putative reference genes. After validation by RT-qPCR, miR-101, miR-125a-5p, miR-148b, miR-151-3p, miR-151-5p, miR-181a, miR-181b miR-29c, miR-324-3p, miR-424, miR-874, RNU6B, RNU48, and Z30 were used for geNorm, NormFinder, and BestKeeper analyses that gave different combinations of recommended reference genes for normalization. Conclusions: The present study provided the first systematic analysis for identifying suitable reference miRNAs for miRNA expression studies of urothelial carcinoma by RT-qPCR. Different combinations of reference genes resulted in reliable expression data for both strongly and less strongly altered miRNAs. Notably, RNU6B, which is the most frequently used reference gene for miRNA studies, gave inaccurate normalization. The combination of four (miR-125a-5p, miR-148b, miR-151-3p, and miR-151-5p) or three (miR-148b, miR-874, miR-181b) miRNA reference genes is recommended for normalization. Candidate reference miRNAs (for RT-qPCR analysis) were selected from 24 papillary urothelial carcinoma and normal bladder tissue samples by miRNA microarrays.

OTHER RELATED OMICS DATASETS IN: PRJNA152929

ORGANISM(S): Homo Sapiens

TISSUE(S): High-grade Papillary Urothelial Carcinoma, Nonmaligant Bladder, Low-grade Papillary Urothelial Carcinoma

SUBMITTER: K Miller   Hellmuth-A Meyer  E Kilic  H A Meyer  K Jung  N Ratert  A Erbersdobler  M Jung  H J Mollenkopf  S Weikert  I Wagner 

PROVIDER: E-GEOD-36121 | ArrayExpress | 2013-02-26

SECONDARY ACCESSION(S): GSE36121PRJNA152929

REPOSITORIES: GEO, ArrayExpress

Dataset's files

Source:
Action DRS
E-GEOD-36121.idf.txt Idf
E-GEOD-36121.processed.1.zip Processed
E-GEOD-36121.raw.1.zip Raw
E-GEOD-36121.sdrf.txt Txt
Items per page:
1 - 4 of 4
altmetric image

Publications


<h4>Background/objective</h4>Reverse transcription quantitative real-time PCR (RT-qPCR) is widely used in microRNA (miRNA) expression studies on cancer. To compensate for the analytical variability produced by the multiple steps of the method, relative quantification of the measured miRNAs is required, which is based on normalization to endogenous reference genes. No study has been performed so far on reference miRNAs for normalization of miRNA expression in urothelial carcinoma. The aim of this  ...[more]

Similar Datasets

| E-GEOD-36121 | BioStudies
| S-EPMC3380005 | BioStudies
| S-EPMC6349320 | BioStudies
| S-EPMC4156210 | BioStudies
| S-EPMC5578666 | BioStudies
| S-EPMC4866712 | BioStudies
| S-EPMC4357047 | BioStudies
| S-EPMC7576785 | BioStudies
| S-EPMC6995162 | BioStudies
| S-EPMC6367481 | BioStudies