Reference miRNAs for colorectal cancer: analysis and verification of current data.
AffiliationClinical Biochemistry section, Department of Neurosciences, Biomedicine and Movement Sciences, University of Verona, Verona, Italy.
MetadataShow full item record
AbstractMicroRNAs (miRNAs) hold great promise in cancer research. The use of appropriate reference miRNAs for normalization of qPCR data is crucial for accurate expression analysis. We present here analysis and verification of current data, proposing a workflow strategy for identification of reference miRNAs in colorectal cancer (CRC). We performed a systematic review of studies aimed to identify stable reference miRNAs in CRC through high-throughput screening. Among the candidate miRNAs selected from the literature we excluded those predicted to target oncogenes or tumor suppressor gene. We then assessed the expression levels of the remaining candidates in exosomes, plasma and tissue samples from CRC patients and healthy controls. The expression stability was evaluated by box-plot, ∆Cq analysis, NormFinder and BestKeeper statistical algorithms. The effects of normalisers on the relative quantification of the oncogenic miR-1290 was also assessed. Our results consistently showed that different combinations of miR-520d, miR-1228 and miR-345 provided the most stably expressed reference miRNAs in the three biological matrices. We identified suitable reference miRNAs for future miRNA expression studies in exosomes plasma and tissues CRC samples. We also provided a novel conceptual framework that overcome the need of performing ex novo identification of suitable reference genes in single experimental systems.
CitationReference miRNAs for colorectal cancer: analysis and verification of current data. 2017, 7 (1):8413 Sci Rep
- MicroRNA expression profiling to identify and validate reference genes for relative quantification in colorectal cancer.
- Authors: Chang KH, Mestdagh P, Vandesompele J, Kerin MJ, Miller N
- Issue date: 2010 Apr 29
- MicroRNA Expression Profiling to Identify and Validate Reference Genes for the Relative Quantification of microRNA in Rectal Cancer.
- Authors: Eriksen AH, Andersen RF, Pallisgaard N, Sørensen FB, Jakobsen A, Hansen TF
- Issue date: 2016
- Identification of stably expressed reference small non-coding RNAs for microRNA quantification in high-grade serous ovarian carcinoma tissues.
- Authors: Bignotti E, Calza S, Tassi RA, Zanotti L, Bandiera E, Sartori E, Odicino FE, Ravaggi A, Todeschini P, Romani C
- Issue date: 2016 Dec
- MicroRNA Expression in Formalin-fixed Paraffin-embedded Cancer Tissue: Identifying Reference MicroRNAs and Variability.
- Authors: Boisen MK, Dehlendorff C, Linnemann D, Schultz NA, Jensen BV, Høgdall EV, Johansen JS
- Issue date: 2015 Dec 29
- Identification and validation of reference genes for qPCR detection of serum microRNAs in colorectal adenocarcinoma patients.
- Authors: Zheng G, Wang H, Zhang X, Yang Y, Wang L, Du L, Li W, Li J, Qu A, Liu Y, Wang C
- Issue date: 2013