Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications

We aimed at assessing the stability of candidate reference genes in urine sediments of men subjected to digital rectal examination for suspected prostate cancer (PCa). Two microRNAs (miR-191 and miR-25) and 1 small nucleolar RNA (SNORD48) were assayed in 35 post-DRE urine sediments of men with PCa a...

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Main Authors: Maria Giulia Egidi, Giovanni Cochetti, Gabriella Guelfi, Danilo Zampini, Silvana Diverio, Giulia Poli, Ettore Mearini
Format: Article
Language:English
Published: Hindawi Limited 2015-01-01
Series:Disease Markers
Online Access:http://dx.doi.org/10.1155/2015/973597
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spelling doaj-cc96f49b580a45da9b2fb58a114d22f92020-11-24T22:03:22ZengHindawi LimitedDisease Markers0278-02401875-86302015-01-01201510.1155/2015/973597973597Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA ApplicationsMaria Giulia Egidi0Giovanni Cochetti1Gabriella Guelfi2Danilo Zampini3Silvana Diverio4Giulia Poli5Ettore Mearini6Department of Surgical and Biomedical Sciences, Institution of Urological, Andrological Surgery and Minimally Invasive Techniques, University of Perugia, Loc. S. Andrea delle Fratte, 06156 Perugia, ItalyDepartment of Surgical and Biomedical Sciences, Institution of Urological, Andrological Surgery and Minimally Invasive Techniques, University of Perugia, Loc. S. Andrea delle Fratte, 06156 Perugia, ItalyDepartment of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, ItalyDepartment of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, ItalyDepartment of Veterinary Medicine, University of Perugia, Via San Costanzo 4, 06126 Perugia, ItalyDepartment of Surgical and Biomedical Sciences, Institution of Urological, Andrological Surgery and Minimally Invasive Techniques, University of Perugia, Loc. S. Andrea delle Fratte, 06156 Perugia, ItalyDepartment of Surgical and Biomedical Sciences, Institution of Urological, Andrological Surgery and Minimally Invasive Techniques, University of Perugia, Loc. S. Andrea delle Fratte, 06156 Perugia, ItalyWe aimed at assessing the stability of candidate reference genes in urine sediments of men subjected to digital rectal examination for suspected prostate cancer (PCa). Two microRNAs (miR-191 and miR-25) and 1 small nucleolar RNA (SNORD48) were assayed in 35 post-DRE urine sediments of men with PCa and in 26 subjects with histologically confirmed benign prostatic hyperplasia (BPH). The stability of candidate reference genes was assessed through BestKeeper algorithm and equivalence test. miR-200b and miR-452 were used to test for the effect of normalization on target genes. Our results proved miR-191 to be the most stable gene, showing the lowest degree of variation and the highest stability value. miR-25 and SNORD48 values fell beyond the cutoff of acceptability. In conclusion, we recommend the use of miR-191 for normalization purposes in post-DRE urine sediments.http://dx.doi.org/10.1155/2015/973597
collection DOAJ
language English
format Article
sources DOAJ
author Maria Giulia Egidi
Giovanni Cochetti
Gabriella Guelfi
Danilo Zampini
Silvana Diverio
Giulia Poli
Ettore Mearini
spellingShingle Maria Giulia Egidi
Giovanni Cochetti
Gabriella Guelfi
Danilo Zampini
Silvana Diverio
Giulia Poli
Ettore Mearini
Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications
Disease Markers
author_facet Maria Giulia Egidi
Giovanni Cochetti
Gabriella Guelfi
Danilo Zampini
Silvana Diverio
Giulia Poli
Ettore Mearini
author_sort Maria Giulia Egidi
title Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications
title_short Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications
title_full Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications
title_fullStr Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications
title_full_unstemmed Stability Assessment of Candidate Reference Genes in Urine Sediment of Prostate Cancer Patients for miRNA Applications
title_sort stability assessment of candidate reference genes in urine sediment of prostate cancer patients for mirna applications
publisher Hindawi Limited
series Disease Markers
issn 0278-0240
1875-8630
publishDate 2015-01-01
description We aimed at assessing the stability of candidate reference genes in urine sediments of men subjected to digital rectal examination for suspected prostate cancer (PCa). Two microRNAs (miR-191 and miR-25) and 1 small nucleolar RNA (SNORD48) were assayed in 35 post-DRE urine sediments of men with PCa and in 26 subjects with histologically confirmed benign prostatic hyperplasia (BPH). The stability of candidate reference genes was assessed through BestKeeper algorithm and equivalence test. miR-200b and miR-452 were used to test for the effect of normalization on target genes. Our results proved miR-191 to be the most stable gene, showing the lowest degree of variation and the highest stability value. miR-25 and SNORD48 values fell beyond the cutoff of acceptability. In conclusion, we recommend the use of miR-191 for normalization purposes in post-DRE urine sediments.
url http://dx.doi.org/10.1155/2015/973597
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