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Identification of reference genes for circulating microRNA analysis in colorectal cancer
Quantitative real-time PCR (qPCR) is the most frequently used method for measuring expression levels of microRNAs (miRNAs), which is based on normalization to endogenous references. Although circulating miRNAs have been regarded as potential non-invasive biomarker of disease, no study has been perfo...
Autores principales: | , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Nature Publishing Group
2016
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5069661/ https://www.ncbi.nlm.nih.gov/pubmed/27759076 http://dx.doi.org/10.1038/srep35611 |
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author | Niu, Yanqin Wu, Yike Huang, Jinyong Li, Qing Kang, Kang Qu, Junle Li, Furong Gou, Deming |
author_facet | Niu, Yanqin Wu, Yike Huang, Jinyong Li, Qing Kang, Kang Qu, Junle Li, Furong Gou, Deming |
author_sort | Niu, Yanqin |
collection | PubMed |
description | Quantitative real-time PCR (qPCR) is the most frequently used method for measuring expression levels of microRNAs (miRNAs), which is based on normalization to endogenous references. Although circulating miRNAs have been regarded as potential non-invasive biomarker of disease, no study has been performed so far on reference miRNAs for normalization in colorectal cancer. In this study we tried to identify optimal reference miRNAs for qPCR analysis across colorectal cancer patients and healthy individuals. 485 blood-derived miRNAs were profiled in serum sample pools of both colorectal cancer and healthy control. Seven candidate miRNAs chosen from profiling results as well as three previous reported reference miRNAs were validated using qPCR in 30 colorectal cancer patients and 30 healthy individuals, and thereafter analyzed by statistical algorithms BestKeeper, geNorm and NormFinder. Taken together, hsa-miR-93-5p, hsa-miR-25-3p and hsa-miR-106b-5p were recommended as a set of suitable reference genes. More interestingly, the three miRNAs validated from 485 miRNAs are derived from a single primary transcript, indicting the cluster may be highly conserved in colorectal cancer. However, all three miRNAs differed significantly between healthy individuals and non-small cell lung cancer or breast cancer patients and could not be used as reference genes in the two types of cancer. |
format | Online Article Text |
id | pubmed-5069661 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2016 |
publisher | Nature Publishing Group |
record_format | MEDLINE/PubMed |
spelling | pubmed-50696612016-10-26 Identification of reference genes for circulating microRNA analysis in colorectal cancer Niu, Yanqin Wu, Yike Huang, Jinyong Li, Qing Kang, Kang Qu, Junle Li, Furong Gou, Deming Sci Rep Article Quantitative real-time PCR (qPCR) is the most frequently used method for measuring expression levels of microRNAs (miRNAs), which is based on normalization to endogenous references. Although circulating miRNAs have been regarded as potential non-invasive biomarker of disease, no study has been performed so far on reference miRNAs for normalization in colorectal cancer. In this study we tried to identify optimal reference miRNAs for qPCR analysis across colorectal cancer patients and healthy individuals. 485 blood-derived miRNAs were profiled in serum sample pools of both colorectal cancer and healthy control. Seven candidate miRNAs chosen from profiling results as well as three previous reported reference miRNAs were validated using qPCR in 30 colorectal cancer patients and 30 healthy individuals, and thereafter analyzed by statistical algorithms BestKeeper, geNorm and NormFinder. Taken together, hsa-miR-93-5p, hsa-miR-25-3p and hsa-miR-106b-5p were recommended as a set of suitable reference genes. More interestingly, the three miRNAs validated from 485 miRNAs are derived from a single primary transcript, indicting the cluster may be highly conserved in colorectal cancer. However, all three miRNAs differed significantly between healthy individuals and non-small cell lung cancer or breast cancer patients and could not be used as reference genes in the two types of cancer. Nature Publishing Group 2016-10-19 /pmc/articles/PMC5069661/ /pubmed/27759076 http://dx.doi.org/10.1038/srep35611 Text en Copyright © 2016, The Author(s) http://creativecommons.org/licenses/by/4.0/ This work is licensed under a Creative Commons Attribution 4.0 International License. The images or other third party material in this article are included in the article’s Creative Commons license, unless indicated otherwise in the credit line; if the material is not included under the Creative Commons license, users will need to obtain permission from the license holder to reproduce the material. To view a copy of this license, visit http://creativecommons.org/licenses/by/4.0/ |
spellingShingle | Article Niu, Yanqin Wu, Yike Huang, Jinyong Li, Qing Kang, Kang Qu, Junle Li, Furong Gou, Deming Identification of reference genes for circulating microRNA analysis in colorectal cancer |
title | Identification of reference genes for circulating microRNA analysis in colorectal cancer |
title_full | Identification of reference genes for circulating microRNA analysis in colorectal cancer |
title_fullStr | Identification of reference genes for circulating microRNA analysis in colorectal cancer |
title_full_unstemmed | Identification of reference genes for circulating microRNA analysis in colorectal cancer |
title_short | Identification of reference genes for circulating microRNA analysis in colorectal cancer |
title_sort | identification of reference genes for circulating microrna analysis in colorectal cancer |
topic | Article |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC5069661/ https://www.ncbi.nlm.nih.gov/pubmed/27759076 http://dx.doi.org/10.1038/srep35611 |
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