ArticlePloS one2016
MicroRNA Expression Profiling to Identify and Validate Reference Genes for the Relative Quantification of microRNA in Rectal Cancer.
Article in PloS one, 2016. The graph could read no effect estimate from its abstract, so it casts no vote on the map. Cited by 17 papers, 1 of them a synthesis that pooled it.
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17 citing papers in PubMed, 1 synthesis or guideline pooled it, 34 citations in OpenAlex.
- Reference miRNAs for colorectal cancer: analysis and verification of current data.Scientific reports · 2017Pooled it
- Tumour budding and poorly differentiated clusters in colon cancer - different manifestations of partial epithelial-mesenchymal transition.The Journal of pathology · 2022Article
- Circulating MicroRNA Panel as a Diagnostic Marker for Hepatocellular Carcinoma.The Turkish journal of gastroenterology : the official journal of Turkish Society of Gastroenterology · 2022Article
- Reference Genes for qPCR-Based miRNA Expression Profiling in 14 Human Tissues.Medical principles and practice : international journal of the Kuwait University, Health Science Centre · 2022Review
- Review
- Urinary Exosomal MicroRNA Signatures in Nephrotic, Biopsy-Proven Diabetic Nephropathy.Journal of clinical medicine · 2020Article
- A systemic approach to screening high-throughput RT-qPCR data for a suitable set of reference circulating miRNAs.BMC genomics · 2020Article
- MicroRNA: Another Pharmacological Avenue for Colorectal Cancer?Frontiers in cell and developmental biology · 2020Review
- MicroRNA expression studies: challenge of selecting reliable reference controls for data normalization.Cellular and molecular life sciences : CMLS · 2019Review
- Identification of suitable reference microRNA for qPCR analysis in pediatric inflammatory bowel disease.Physiological genomics · 2019Article
- Functional Effects ofCancers · 2019Article
- Toxicity Evaluation and Biomarker Selection with Validated Reference Gene in Embryonic Zebrafish Exposed to Mitoxantrone.International journal of molecular sciences · 2018Article
- Endothelial Cell Aging: How miRNAs Contribute?Journal of clinical medicine · 2018Review
- Differential plasma microvesicle and brain profiles of microRNA in experimental cerebral malaria.Malaria journal · 2018Article
- Association between the expression of microRNAs and the response of patients with locally advanced rectal cancer to preoperative chemoradiotherapy.Oncology letters · 2017Article
- Validation of optimal reference genes for quantitative real time PCR in muscle and adipose tissue for obesity and diabetes research.Scientific reports · 2017Article
- Article
Corrections and comments
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Authors and funding
6 authors at 3 institutions in 1 country.
Funding
No grant is acknowledged in the PubMed record.
Abstract
introductionMicroRNAs (miRNAs) play important roles in regulating biological processes at the post-transcriptional level. Deregulation of miRNAs has been observed in cancer, and miRNAs are being investigated as potential biomarkers regarding diagnosis, prognosis and prediction in cancer management. Real-time quantitative polymerase chain reaction (RT-qPCR) is commonly used, when measuring miRNA expression. Appropriate normalisation of RT-qPCR data is important to ensure reliable results. The aim of the present study was to identify stably expressed miRNAs applicable as normaliser candidates in future studies of miRNA expression in rectal cancer. MATERIALS AND
methodsWe performed high-throughput miRNA profiling (OpenArray®) on ten pairs of laser micro-dissected rectal cancer tissue and adjacent stroma. A global mean expression normalisation strategy was applied to identify the most stably expressed miRNAs for subsequent validation. In the first validation experiment, a panel of miRNAs were analysed on 25 pairs of micro dissected rectal cancer tissue and adjacent stroma. Subsequently, the same miRNAs were analysed in 28 pairs of rectal cancer tissue and normal rectal mucosa.
resultsFrom the miRNA profiling experiment, miR-645, miR-193a-5p, miR-27a and let-7g were identified as stably expressed, both in malignant and stromal tissue. In addition, NormFinder confirmed high expression stability for the four miRNAs. In the RT-qPCR based validation experiments, no significant difference between tumour and stroma/normal rectal mucosa was detected for the mean of the normaliser candidates miR-27a, miR-193a-5p and let-7g (first validation P = 0.801, second validation P = 0.321). MiR-645 was excluded from the data analysis, because it was undetected in 35 of 50 samples (first validation) and in 24 of 56 samples (second validation), respectively. Significant difference in expression level of RNU6B was observed between tumour and adjacent stromal (first validation), and between tumour and normal rectal mucosa (second validation).
conclusionWe recommend the mean expression of miR-27a, miR-193a-5p and let-7g as normalisation factor, when performing miRNA expression analyses by RT-qPCR on rectal cancer tissue.
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