Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of
Objective: In spite of accumulating information about pathological aspects of sulfur mustard (SM), the precise mechanism responsible for its effects is not well understood. Circulating microRNAs (miRNAs) are promising biomarkers for disease diagnosis and prognosis. Accurate normalization using ap...
| Main Authors: | , , , , , , , |
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| Format: | Article |
| Language: | English |
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Royan Institute (ACECR), Tehran
2015-10-01
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| Series: | Cell Journal |
| Subjects: | |
| Online Access: | http://celljournal.org/web/journal/article/979/download |
| _version_ | 1818048780300189696 |
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| author | Sedigheh Gharbi Mehdi Shamsara Shahriar Khateri Mohammad Reza Soroush Nassim Ghorbanmehr Mahmood Tavallaei Mohammad Reza Nourani Seyed Javad Mowla |
| author_facet | Sedigheh Gharbi Mehdi Shamsara Shahriar Khateri Mohammad Reza Soroush Nassim Ghorbanmehr Mahmood Tavallaei Mohammad Reza Nourani Seyed Javad Mowla |
| author_sort | Sedigheh Gharbi |
| collection | DOAJ |
| description | Objective: In spite of accumulating information about pathological aspects of sulfur
mustard (SM), the precise mechanism responsible for its effects is not well understood.
Circulating microRNAs (miRNAs) are promising biomarkers for disease diagnosis and
prognosis. Accurate normalization using appropriate reference genes, is a critical step in
miRNA expression studies. In this study, we aimed to identify appropriate reference gene
for microRNA quantification in serum samples of SM victims.
Materials and Methods: In this case and control experimental study, using quantitative
real-time polymerase chain reaction (qRT-PCR), we evaluated the suitability of a panel
of small RNAs including SNORD38B, SNORD49A, U6, 5S rRNA, miR-423-3p, miR-191,
miR-16 and miR-103 in sera of 28 SM-exposed veterans of Iran-Iraq war (1980-1988)
and 15 matched control volunteers. Different statistical algorithms including geNorm,
Normfinder, best-keeper and comparative delta-quantification cycle (Cq) method were
employed to find the least variable reference gene.
Results: miR-423-3p was identified as the most stably expressed reference gene, and miR-
103 and miR-16 ranked after that.
Conclusion: We demonstrate that non-miRNA reference genes have the least stability
in serum samples and that some house-keeping miRNAs may be used as more
reliable reference genes for miRNAs in serum. In addition, using the geometric mean
of two reference genes could increase the reliability of the normalizers. |
| first_indexed | 2024-12-10T10:27:07Z |
| format | Article |
| id | doaj.art-c107b5d227a04b54808d3b25dcb1b3d4 |
| institution | Directory Open Access Journal |
| issn | 2228-5806 2228-5814 |
| language | English |
| last_indexed | 2024-12-10T10:27:07Z |
| publishDate | 2015-10-01 |
| publisher | Royan Institute (ACECR), Tehran |
| record_format | Article |
| series | Cell Journal |
| spelling | doaj.art-c107b5d227a04b54808d3b25dcb1b3d42022-12-22T01:52:41ZengRoyan Institute (ACECR), TehranCell Journal2228-58062228-58142015-10-01173494501Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples ofSedigheh Gharbi0Mehdi Shamsara1Shahriar Khateri2Mohammad Reza Soroush3Nassim Ghorbanmehr4Mahmood Tavallaei5Mohammad Reza Nourani6Seyed Javad Mowla7Department of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranNational Institute of Genetic Engineering and Biotechnology, Tehran, IranJanbazan Medical and Engineering Research Center (JMERC), Tehran, IranJanbazan Medical and Engineering Research Center (JMERC), Tehran, IranDepartmen of Biotechnology, Faculty of Biological Sciences, Alzahra University, Tehran, IranGenetic Research Center, Baqiyatallah University of Medical Sciences, Tehran, IranChemical Injury Research Center (CIRC), Baqiyatallah University of Medical Sciences, Tehran, IranDepartment of Molecular Genetics, Faculty of Biological Sciences, Tarbiat Modares University, Tehran, IranObjective: In spite of accumulating information about pathological aspects of sulfur mustard (SM), the precise mechanism responsible for its effects is not well understood. Circulating microRNAs (miRNAs) are promising biomarkers for disease diagnosis and prognosis. Accurate normalization using appropriate reference genes, is a critical step in miRNA expression studies. In this study, we aimed to identify appropriate reference gene for microRNA quantification in serum samples of SM victims. Materials and Methods: In this case and control experimental study, using quantitative real-time polymerase chain reaction (qRT-PCR), we evaluated the suitability of a panel of small RNAs including SNORD38B, SNORD49A, U6, 5S rRNA, miR-423-3p, miR-191, miR-16 and miR-103 in sera of 28 SM-exposed veterans of Iran-Iraq war (1980-1988) and 15 matched control volunteers. Different statistical algorithms including geNorm, Normfinder, best-keeper and comparative delta-quantification cycle (Cq) method were employed to find the least variable reference gene. Results: miR-423-3p was identified as the most stably expressed reference gene, and miR- 103 and miR-16 ranked after that. Conclusion: We demonstrate that non-miRNA reference genes have the least stability in serum samples and that some house-keeping miRNAs may be used as more reliable reference genes for miRNAs in serum. In addition, using the geometric mean of two reference genes could increase the reliability of the normalizers.http://celljournal.org/web/journal/article/979/downloadMicroRNAQuantitative Real Time-PCRNormalizationSulfur MustardmiR-423 |
| spellingShingle | Sedigheh Gharbi Mehdi Shamsara Shahriar Khateri Mohammad Reza Soroush Nassim Ghorbanmehr Mahmood Tavallaei Mohammad Reza Nourani Seyed Javad Mowla Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of Cell Journal MicroRNA Quantitative Real Time-PCR Normalization Sulfur Mustard miR-423 |
| title | Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of |
| title_full | Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of |
| title_fullStr | Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of |
| title_full_unstemmed | Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of |
| title_short | Identification of Reliable Reference Genes for Quantification of MicroRNAs in Serum Samples of |
| title_sort | identification of reliable reference genes for quantification of micrornas in serum samples of |
| topic | MicroRNA Quantitative Real Time-PCR Normalization Sulfur Mustard miR-423 |
| url | http://celljournal.org/web/journal/article/979/download |
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