Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR

Marabita, Francesco; Candia, Paola de; Torri, A; Tegnér, Jesper; Abrignani, Sergio; Rossi, Riccardo L.

doi:10.1093/bib/bbv056

Cited by 217 publications

(221 citation statements)

References 56 publications

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“…and miR-1, miR-206 and miR-133b identified by Koutsoulidou and collaborators24. Normalization is a critical issue in all circulating miRNA studies26. Three normalizers were used, one exogenous (cel-miR-39) and two endogenous (miR-106a and miR-17-5p).…”

Section: Resultsmentioning

confidence: 99%

Validation of plasma microRNAs as biomarkers for myotonic dystrophy type 1

Perfetti

Greco

Cardani

et al. 2016

Sci Rep

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Non-invasive and simple to measure biomarkers are still an unmet need for myotonic dystrophy type 1 (DM1). Indeed, muscle biopsies can be extremely informative, but their invasive nature limits their application. Extracellular microRNAs are emerging humoral biomarkers and preliminary studies identified a group of miRNAs that are deregulated in the plasma or serum of small groups of DM1 patients. Here we adopted very stringent selection and normalization criteria to validate or disprove these miRNAs in 103 DM1 patients and 111 matched controls. We confirmed that 8 miRNAs out of 12 were significantly deregulated in DM1 patients: miR-1, miR-27b, miR-133a, miR-133b, miR-206, miR-140-3p, miR-454 and miR-574. The levels of these miRNAs, alone or in combination, discriminated DM1 from controls significantly, and correlated with both skeletal muscle strength and creatine kinase values. Interestingly, miR-133b levels were significantly higher in DM1 female patients. Finally, the identified miRNAs were also deregulated in the plasma of a small group (n = 30) of DM2 patients. In conclusion, this study proposes that miRNAs might be useful as DM1 humoral biomarkers.

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Section: Resultsmentioning

confidence: 99%

Validation of plasma microRNAs as biomarkers for myotonic dystrophy type 1

Perfetti

Greco

Cardani

et al. 2016

Sci Rep

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“…Indeed, we did not use miR-16, as in many studies conducted by others, since this miRNA is particularly susceptible to hemolysis. 25 In this pilot study, we identified five miRNAs with promising diagnostic power in plasma of melanoma patients at different stages of disease. Three of them, that is, miR-149-3p, miR-150-5p, and miR-15b-5p, were significantly up-regulated, while two, that is, miR-193a-3p and miR-524-5p, were significantly down-regulated in plasma of patients compared with healthy controls.…”

Section: Discussionmentioning

confidence: 99%

Identification of plasma microRNAs as new potential biomarkers with high diagnostic power in human cutaneous melanoma

et al. 2017

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Melanoma is a devastating disease with few therapeutic options in the advanced stage and with the urgent need of reliable biomarkers for early detection. In this context, circulating microRNAs are raising great interest as diagnostic biomarkers. We analyzed the expression profiles of 21 selected microRNAs in plasma samples from melanoma patients and healthy donors to identify potential diagnostic biomarkers. Data analysis was performed using global mean normalization and NormFinder algorithm. Linear regression followed by receiver operating characteristic analyses was carried out to evaluate whether selected plasma miRNAs were able to discriminate between cases and controls. We found five microRNAs that were differently expressed among cases and controls after Bonferroni correction for multiple testing. Specifically, miR-15b-5p, miR-149-3p, and miR-150-5p were up-regulated in plasma of melanoma patients compared with healthy controls, while miR-193a-3p and miR-524-5p were down-regulated. Receiver operating characteristic analyses of these selected microRNAs provided area under the receiver operating characteristic curve values ranging from 0.80 to 0.95. Diagnostic value of microRNAs is improved when considering the combination of miR-149-3p, miR-150-5p, and miR-193a-3p. The triple classifier had a high capacity to discriminate between melanoma patients and healthy controls, making it suitable to be used in early melanoma diagnosis.

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“…External reference couldn’t correct for the other causes of variability such as the total concentration of miRNA fraction. Endogenous controls were required19. To our knowledge, no study has been involved in the evaluation of reference genes specific for serum miRNAs in osteoporosis.…”

mentioning

confidence: 99%

Identification of suitable reference gene and biomarkers of serum miRNAs for osteoporosis

Chen

Pang

et al. 2016

Sci Rep

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Our objective was to identify suitable reference genes in serum miRNA for normalization and screen potential new biomarkers for osteoporosis diagnosis by a systematic study. Two types of osteoporosis models were used like as mechanical unloading and estrogen deficiency. Through a large-scale screening using microarray, qPCR validation and statistical algorithms, we first identified miR-25-3p as a suitable reference gene for both type of osteoporosis, which also showed stability during the differentiation processes of osteoblast and osteoclast. Then 15 serum miRNAs with differential expression in OVX rats were identified by microarray and qPCR validation. We further detected these 15 miRNAs in postmenopausal women and bedrest rhesus monkeys and evaluated their diagnostic value by ROC analysis. Among these miRNAs, miR-30b-5p was significantly down-regulated in postmenopausal women with osteopenia or osteoporosis; miR-103-3p, miR-142-3p, miR-328-3p were only significantly decreased in osteoporosis. They all showed positive correlations with BMD. Except miR328-3p, the other three miRNAs were also declined in the rhesus monkeys after long-duration bedrest. Their AUC values (all >0.75) proved the diagnostic potential. Our results provided a reliable normalization reference gene and verified a group of circulating miRNAs as non-invasive biomarkers in the detection of postmenopausal- and mechanical unloading- osteoporosis.

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Normalization of circulating microRNA expression data obtained by quantitative real-time RT-PCR

Cited by 217 publications

References 56 publications

Validation of plasma microRNAs as biomarkers for myotonic dystrophy type 1

Validation of plasma microRNAs as biomarkers for myotonic dystrophy type 1

Identification of plasma microRNAs as new potential biomarkers with high diagnostic power in human cutaneous melanoma

Identification of suitable reference gene and biomarkers of serum miRNAs for osteoporosis

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