Validation of suitable endogenous control genes for expression studies of miRNA in prostate cancer tissues

Carlsson, Jessica; Helenius, Gisela; Karlsson, Mats G.; Lubovac, Zelmina; Andrén, Ove; Olsson, Björn; Klinga-Levan, Karin

doi:10.1016/j.cancergencyto.2010.06.009

Cited by 30 publications

(22 citation statements)

References 15 publications

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“…On the other hand, Schaefer et al [40] recommended the use of miR-130b or the geometric mean of miR-130b and RNU6-2 to normalize miRNA expression in PCa tissues studies. However, according to Carlsson et al [41], RNU24 and RNU44 would be suitable control genes for miRNA expression studies in prostate tissues, but not miR-130b, because its stability was not appropriate to be a normalizer control.…”

Section: Preanalytical and Analytical Variables In Mirna Measurementmentioning

confidence: 99%

miRNAs as novel biomarkers in the management of prostate cancer

Filella

Foj

2017

Clinical Chemistry and Laboratory Medicine (CCLM)

100

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microRNAs (miRNAs) are small non-coding RNAs that control gene expression posttranscriptionally and are part of the giant non codifying genoma. Cumulating data suggest that miRNAs are promising potential biomarkers for many diseases, including cancer. Prostate cancer (PCa) detection is currently based in the serum prostate-specific antigen biomarker and digital rectal examination. However, these methods are limited by a low predictive value and the adverse consequences associated with overdiagnosis and overtreatment. New biomarkers that could be used for PCa detection and prognosis are still needed. Recent studies have demonstrated that aberrant expressions of microRNAs are associated with the underlying mechanisms of PCa. This review attempts to extensively summarize the current knowledge of miRNA expression patterns, as well as their targets and involvement in PCa pathogenesis. We focused our review in the value of circulating and urine miRNAs as biomarkers in PCa patients, highlighting the existing discrepancies between different studies, probably associated with the important methodological issues related to their quantitation and normalization. The majority of studies have been performed in serum or plasma, but urine obtained after prostate massage appears as a new way to explore the usefulness of miRNAs. Large screening studies to select a miRNA profile have been completed, but bioinformatics tools appear as a new approach to select miRNAs that are relevant in PCa development.Promising preliminary results were published concerning miR-141, miR-375 and miR-21, but larger and prospective studies using standardized methodology are necessary to define the value of miRNAs in the detection and prognosis of PCa.

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Section: Preanalytical and Analytical Variables In Mirna Measurementmentioning

confidence: 99%

miRNAs as novel biomarkers in the management of prostate cancer

Filella

Foj

2017

Clinical Chemistry and Laboratory Medicine (CCLM)

100

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“…Relative expression levels were calculated using the comparative Ct method with the small nuclear RNA RNU24 used as an endogenous quantitative normalisation control as it has been identified as being the most consistent option with the least variability for prostate cancer (Carlsson et al 2010). …”

Section: Rt-qpcr Analysis To Assess Mirna Expressionmentioning

confidence: 99%

A genetic variant of MDM4 influences regulation by multiple microRNAs in prostate cancer

Stegeman¹,

Moya²,

Selth³

et al. 2015

Endocrine-Related Cancer

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The oncogene MDM4, also known as MDMX or HDMX, contributes to cancer susceptibility and progression through its capacity to negatively regulate a range of genes with tumoursuppressive functions. As part of a recent genome-wide association study it was determined that the A-allele of the rs4245739 SNP (AOC), located in the 3 0 -UTR of MDM4, is associated with an increased risk of prostate cancer. Computational predictions revealed that the rs4245739 SNP is located within a predicted binding site for three microRNAs (miRNAs): miR-191-5p, miR-887 and miR-3669. Herein, we show using reporter gene assays and endogenous MDM4 expression analyses that miR-191-5p and miR-887 have a specific affinity for the rs4245739 SNP C-allele in prostate cancer. These miRNAs do not affect MDM4 mRNA levels, rather they inhibit its translation in C-allele-containing PC3 cells but not in LNCaP cells homozygous for the A-allele. By analysing gene expression datasets from patient cohorts, we found that MDM4 is associated with metastasis and prostate cancer progression and that targeting this gene with miR-191-5p or miR-887 decreases in PC3 cell viability. This study is the first, to our knowledge, to demonstrate regulation of the MDM4 rs4245739 SNP C-allele by two miRNAs in prostate cancer, and thereby to identify a mechanism by which the MDM4 rs4245739 SNP A-allele may be associated with an increased risk for prostate cancer.Key Words " MDM4" microRNA " single nucleotide polymorphism " prostate cancer

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“…Similarly, the stability of both SNORD48 and miR-16 following irradiation was high in all cell lines tested. SNORD48 was previously reported as a stable endogenous control for miRNA studies in prostate cancer tissues with a stability value of 2.05 using NormFinder [28].…”

Section: Discussionmentioning

confidence: 99%

“…A few studies have validated the use of appropriate endogenous controls in prostate cancer [28,29], but the suitability of commonly used endogenous controls in radiation studies has not been investigated. The aim of this study was to evaluate the stability of endogenous controls commonly used in mRNA and miRNA expression studies in irradiated prostate cells.…”

Section: Introductionmentioning

confidence: 99%

Identification of suitable endogenous controls for gene and miRNA expression studies in irradiated prostate cancer cells

et al. 2015

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This study aimed to to evaluate the stability of commonly used endogenous control genes for messenger RNA (mRNA) (N = 16) and miRNAs (N = 3) expression studies in prostate cell lines following irradiation. The stability of endogenous control genes expression in irradiated (6 Gy) versus unirradiated controls was quantified using NormFinder and coefficient of variation analyses. HPRT1 and 18S were identified as most and least stable endogenous controls, respectively, for mRNA expression studies in irradiated prostate cells. SNORD48 and miR16 miRNA endogenous controls tested were associated with low coefficient of variations following irradiation (6 Gy). This study highlights that commonly used endogenous controls can be responsive to radiation and validation is required prior to gene/miRNAs expression studies.

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Validation of suitable endogenous control genes for expression studies of miRNA in prostate cancer tissues

Cited by 30 publications

References 15 publications

miRNAs as novel biomarkers in the management of prostate cancer

miRNAs as novel biomarkers in the management of prostate cancer

A genetic variant of MDM4 influences regulation by multiple microRNAs in prostate cancer

Identification of suitable endogenous controls for gene and miRNA expression studies in irradiated prostate cancer cells

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