Selection of reference genes is critical for miRNA expression analysis in human cardiac tissue. A focus on atrial fibrillation

Masè, Michela; Grasso, Margherita; Avogaro, Laura; D’Amato, Elvira; Tessarolo, Francesco; Graffigna, Angelo; Denti, Michela Alessandra; Ravelli, Flavia

doi:10.1038/srep41127

Cited by 72 publications

(57 citation statements)

References 47 publications

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“…The primers for miRNAs (hsa-miR-15a-5p, hsa-miR-29b-3p, hsa-miR-124-3p, hsa-miR-140-5p, hsa-miR-146a-5p, hsa-miR-148a-3p, hsa-miR-155-5p, hsa-miR-223-3p, hsa-miR-301b, hsa-miR-628-5p) are listed in Table 2. The expression of each miRNA was normalized to the recommended housekeeping genes, human small nucleolar RNA SNORD48 [46] and hsa-miR-16-5p [47]. CT values >35 were excluded.…”

Section: Rna Extraction and Mirna Expression By Rt-qpcrmentioning

confidence: 99%

Salivary Small Extracellular Vesicles Associated miRNAs in Periodontal Status—A Pilot Study

Han

Bartold

Salomón

et al. 2020

IJMS

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This pilot study aims to investigate whether salivary small extracellular vesicle (sEV)-associated microRNAs could act as potential biomarkers for periodontal disease status. Twenty-nine participants (10 who were healthy, nine with gingivitis, 10 with stage III/IV periodontitis) were recruited and unstimulated whole saliva samples were collected. Salivary sEVs were isolated using the size-exclusion chromatography (SEC) method and characterised by morphology, EV-protein and size distribution using transmission electron microscopy (TEM), Western Blot and Nanoparticle Tracking Analysis (NTA), respectively. Ten mature microRNAs (miRNAs) in salivary sEVs and saliva were evaluated using RT-qPCR. The discriminatory power of miRNAs as biomarkers in gingivitis and periodontitis versus healthy controls was evaluated by Receiver Operating Characteristics (ROC) curves. Salivary sEVs were comparable to sEVs morphology, mode, size distribution and particle concentration in healthy, gingivitis and periodontitis patients. Compared to miRNAs in whole saliva, three significantly increased miRNAs (hsa-miR-140-5p, hsa-miR-146a-5p and hsa-miR-628-5p) were only detected in sEVs in periodontitis when compared to that of healthy controls, with a good discriminatory power (area under the curve (AUC) = 0.96) for periodontitis diagnosis. Our study demonstrated that salivary sEVs are a non-invasive source of miRNAs for periodontitis diagnosis. Three miRNAs that are selectively enriched in sEVs, but not whole saliva, could be potential biomarkers for periodontal disease status.

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Section: Rna Extraction and Mirna Expression By Rt-qpcrmentioning

confidence: 99%

Salivary Small Extracellular Vesicles Associated miRNAs in Periodontal Status—A Pilot Study

Han

Bartold

Salomón

et al. 2020

IJMS

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“…PCR was performed and the target genes were normalised to three chosen reference genes. SNORD68, SNORD96A and RNU6-6P were employed as the reference genes to analyse miRNAs expression in the rat cardiac tissues [42][43][44] . The ratio between the target gene and the geometric mean of the reference genes were calculated to achieve mean normalised expression (MNE) of the target genes in each sample.…”

Section: Main Textmentioning

confidence: 99%

The impacts of intrauterine Bisphenol A exposure on pregnancy and expression of miRNAs related to heart development and diseases in animal model

Rasdi

Kamaludin

Ab‐Rahim

et al. 2020

Sci Rep

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this study aimed to examine the impact of BpA exposure on pregnancy and foetuses on cardiac tissues and the expression of cardiac microRnAs (miRnAs) related to heart development and diseases. Pregnancy is known to be the "critical windows" in determining the offspring physical and cells development in their life after birth. The increment of the risk of cardiovascular disease (CVD) in a later stage of life has been reported by few studies demonstrated from prenatal exposure of BpA. BpA has been shown to alter miRNAs expression profiles for organ development, regeneration and metabolic functions. These alterations have been associated with the risk of CVDs. However, the associations between pregnancy outcomes and miRnAs expression in cardiac of mother-and foetuses-exposed to BPA are still not entirely explored. In BPA-exposed pregnant rat groups, a significant weight gained was observed in comparison to control (p < 0.05). Interestingly, significant changes in systolic and diastolic blood pressure between the first and third trimester of BPA-exposed pregnant rats were also observed (p < 0.05). In BPA-exposed pregnant rats, miR-499-5p was significantly altered in the heart (p < 0.01). Meanwhile, altered miR-17-5p,-208-3p, and-210-3p expressions were observed in all heart of the foetuses from BpA-exposed pregnant rats (p < 0.05). In H&E staining, BPA-exposed foetal hearts showed a sign of fibrosis while BPA-exposed pregnant rats showed muscle remnant. Masson trichrome staining further confirmed the presence of fibrosis observed in BPA-exposed foetal heart and reduced expression of cardiac troponin I (cTnI) was also observed in BPA-exposed foetal heart. In summary, altered cardiac miRnAs with histological changes were observed in both mother-and foetus-exposed BPA These findings put forward the importance of future work to further understand how prenatal BPA exposure affect foetuses in their later stage of life. Altered foetal development "programming" may predispose certain individuals to the risk of chronic disease development later in their life. This was suggested by Barker and co-worker who presented the first finding on the increased risk of cardiovascular disease (CVD) in children of malnourished mothers 1. Barker then further extended his theory and linked the CVD development and insulin resistance to the environment of the placenta. His findings have attracted another study to report on the insufficiency of uteroplacental of malnutrition mothers increases the risk of the offspring to type 2 diabetes 2. Another study demonstrated that miR-208, a

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“…U6 and other standard use housekeeping genes have repeatedly proved to be unqualified as internal control in miRNA studies in body fluids. 40,41 Therefore, in this study we used endogenous controls within the same RNA class as recommended. 42 In the screening cohort, both miRNA-17 and miRNA-191 were chosen as internal controls, as they displayed a stable expression level in all samples and had previously proven suitable as controls.…”

Section: Rna Extractionmentioning

confidence: 99%

microRNA expression in tumour tissue and plasma in patients with newly diagnosed metastatic prostate cancer

et al. 2018

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Prostate cancer is the most common cancer among men in the western world. Clinical practice is continuously challenged by the pitfalls of the available diagnostic tools. microRNAs may represent promising biomarkers in many types of human cancers, including prostate cancer. The aim of this study was to investigate microRNA expression in tumour tissue and matched plasma in a cohort of patients with primary metastatic prostate cancer. The relative expression of 12 microRNAs was assessed in diagnostic needle biopsies from the prostate and matched plasma samples in two prospective cohorts (screening cohorts) comprising 21 patients with metastatic prostate cancer and 25 control patients. An independent validation cohort of plasma samples was collected prospectively from 149 newly diagnosed patients with local/locally advanced prostate cancer. Analyses were performed using real-time polymerase chain reaction. miRNA-93 showed a significant negative correlation between expression in tumour tissue and plasma in patients with metastatic prostate cancer. Furthermore, the plasma level of miRNA-93 significantly decreased after treatment in patients with local/locally advanced prostate cancer compared to baseline plasma level. The expression of six microRNAs (let-7b, miRNA-34a, -125b, -143, -145 and -221) was downregulated, and three microRNAs (miRNA-21, -25 and miRNA-93) were upregulated in tumour tissue compared to benign prostate tissue. In plasma, six microRNAs were upregulated (miRNA-21, -125b, -126, -141, -143 and -375), while let-7b was downregulated in patients with metastatic prostate cancer compared to the control cohort. In the metastatic prostate cancer cohort, the expression of four microRNAs (miRNA-125b, -126, -143 and -221), and miRNA-141 in tissue was associated with Gleason score and prostate-specific antigen, respectively. The expression of miRNA-93 in tumour tissue was correlated with matched plasma levels and showed a significant decrease in plasma level after intervention in local prostate cancer. Differential expression between tumour and benign prostate was detected for several microRNAs in both tissue and plasma.

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Selection of reference genes is critical for miRNA expression analysis in human cardiac tissue. A focus on atrial fibrillation

Cited by 72 publications

References 47 publications

Salivary Small Extracellular Vesicles Associated miRNAs in Periodontal Status—A Pilot Study

Salivary Small Extracellular Vesicles Associated miRNAs in Periodontal Status—A Pilot Study

The impacts of intrauterine Bisphenol A exposure on pregnancy and expression of miRNAs related to heart development and diseases in animal model

microRNA expression in tumour tissue and plasma in patients with newly diagnosed metastatic prostate cancer

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