Comparison of miR‐16 and cel‐miR‐39 as reference controls for serum miRNA normalization in colorectal cancer

Madadi, Soheil; Soleimani, Meysam

doi:10.1002/jcb.28174

Cited by 15 publications

(9 citation statements)

References 10 publications

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“…10 Caenorhabditis elegans miR-39 (cel-miR-39) was used as the housekeeping gene for hsa-miR-1275 expression detection. 13 As a result (Figures 2B and 2C), hsa-miR-1275 increased significantly in the serum of EUE compared with normal control, which is consistent with the results of the miRNA microarray. However, hsa-miR-132 exhibited no significant difference in serum.…”

Section: Hsa-mir-1275 Was Frequently Differentially Regulated In Euesupporting

confidence: 86%

“…10 cel-miR-39 was used as the housekeeping gene for the detection of hsa-miR-1275 expression. 13 The primers for hsa-miR-1275, hsa-miR-132, and cel-miR-39 were included in TaqMan miRNA assays (4427975; Applied Biosystems, Life Technologies, NY, USA The U251 cell line (glioblastoma-derived human cell line; Cat. number TCHu 58) was purchased from the cell bank of the Chinese Academy of Sciences (Shanghai, China).…”

Section: Quantitation Of Hsa-mir-1275 By Taqman Assaymentioning

confidence: 99%

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Identification of hsa-miR-1275 as a Novel Biomarker Targeting MECP2 for Human Epilepsy of Unknown Etiology

Zhao

Wang

et al. 2020

Molecular Therapy - Methods & Clinical Development

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Epilepsy affects around 70 million people worldwide, with a 65% rate of unknown etiology. This rate is known as epilepsy of unknown etiology (EUE). Dysregulation of microRNAs (miRNAs) is recognized to contribute to mental disorders, including epilepsy. However, miRNA dysregulation is poorly understood in EUE. Here, we conducted miRNA expression profiling of EUE by microarray technology and identified 57 pathogenic changed miRNAs with significance. The data and bioinformatic analysis results indicated that among these miR-NAs, hsa-microRNA (miR)-1275 was highly associated with neurological disorders. Subsequently, new samples of serum and cerebrospinal fluid were collected for validation of hsa-miR-1275 expression by TaqMan assays. Results show that hsa-miR-1275 in serums of EUE were increased significantly, but in cerebrospinal fluid, the miRNA was decreased. Moreover, the MECP2 gene was selected as a hsa-miR-1275 target based on target prediction tools and gene ontology analysis. Validation of in vitro tests proved that MECP2 expression was specifically inhibited by hsa-miR-1275. Additionally, overexpression of hsa-miR-1275 can elevate expression of nuclear factor kB (NF-kB) and promote cell apoptosis. Taken together, hsa-miR-1275 might represent a novel biomarker targeting MECP2 for human EUE.

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Section: Hsa-mir-1275 Was Frequently Differentially Regulated In Euesupporting

confidence: 86%

Section: Quantitation Of Hsa-mir-1275 By Taqman Assaymentioning

confidence: 99%

Identification of hsa-miR-1275 as a Novel Biomarker Targeting MECP2 for Human Epilepsy of Unknown Etiology

Zhao

Wang

et al. 2020

Molecular Therapy - Methods & Clinical Development

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“…Binderup et al have emphasized the importance of the chosen normalization strategy in RT-qPCR data analyses [303]. The most frequent normalization technique involves the use of an exogenous spike-in of a synthetic miRNA such as cel-miR-39 or an endogenous one like miR-16 as reference genes during the initial phase of RNA extraction [304]. However, it is generally strongly implied that the use of a single reference gene is insufficient for accurate miRNA results, with researchers preferring strategies that apply a combination of control miRNAs, both exogenous and endogenous [305].…”

Section: Limitations Of Circulating Mirnas As Biomarkersmentioning

confidence: 99%

miRNAs as Biomarkers in Disease: Latest Findings Regarding Their Role in Diagnosis and Prognosis

Condrat

Thompson

Barbu

et al. 2020

Cells

843

583

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MicroRNAs (miRNAs) represent a class of small, non-coding RNAs with the main roles of regulating mRNA through its degradation and adjusting protein levels. In recent years, extraordinary progress has been made in terms of identifying the origin and exact functions of miRNA, focusing on their potential use in both the research and the clinical field. This review aims at improving the current understanding of these molecules and their applicability in the medical field. A thorough analysis of the literature consulting resources available in online databases such as NCBI, PubMed, Medline, ScienceDirect, and UpToDate was performed. There is promising evidence that in spite of the lack of standardized protocols regarding the use of miRNAs in current clinical practice, they constitute a reliable tool for future use. These molecules meet most of the required criteria for being an ideal biomarker, such as accessibility, high specificity, and sensitivity. Despite present limitations, miRNAs as biomarkers for various conditions remain an impressive research field. As current techniques evolve, we anticipate that miRNAs will become a routine approach in the development of personalized patient profiles, thus permitting more specific therapeutic interventions.Biogenesis of miRNA begins in the nucleus, where the transcription of its precursor, primary miRNA or pri-miRNA takes place under the influence of RNA polymerases II and III [11,12]. The resulting molecule is a hairpin-like structure, which contains a loop at one end [11]. This primordial mi-RNA precursor that is usually made up of hundreds of nucleotides is then processed consecutively by two RNase III enzymes [13][14][15]. The first enzyme to act upon the pri-miRNA, which still resides in the nucleus, is called Drosha or DCGR8, and turns it into a new hairpin-like structure of approximately 70 nucleotides, the Precursor-miRNA or pre-miRNA. The latter is then transported to the cytoplasm, with the help of Exportin-5, where it is cleaved again by the Ago2/Dicer complex leading to the short, mature miRNA double strands [16].Further on, one of the strands, usually known as the guide strand, will be integrated into the RNA-induced silencing complex (RISC), while the other one, known as the passenger strand, is going to be degraded, even though in some occasions it has been found to be also functional [17]. In most cases, the strand that contains the less stable 5 end or a uracil at the beginning is more likely to be selected as the guide strand [18][19][20]. In those situations, where the passenger strand is not degraded and both get incorporated into the miRISC complex, the mature miRNA in the guide strand will be the dominant one [21,22].The main role of miRNA in the human body is gene regulation [23] by mediating the degradation of mRNA and also by regulating transcription and translation through canonical and non-canonical mechanisms [4]. The canonical mechanism means that the miRISC complex containing the miRNA guide strand is exerting its action by binding to the targ...

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“…The cellular expression of lncRNA-UCA1 and XIAP mRNA was normalized to that of glyceraldehyde 3-phosphate dehydrogenase (GAPDH) 16 . The exosomal level of lncRNA-UCA1 was normalized to that of cel-miR-39 (C39) 17,18 . For quantification of miR-873-5p, cDNA was synthesized with an miRNA First-Strand cDNA Synthesis Kit (by stem-loop) (Vazyme Biotech, China).…”

Section: Quantitative Real-time Pcr (Qrt-pcr)mentioning

confidence: 99%

Long noncoding RNA UCA1 from hypoxia-conditioned hMSC-derived exosomes: a novel molecular target for cardioprotection through miR-873-5p/XIAP axis

et al. 2020

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Exosomes (Exo) secreted from mesenchymal stem cells (hMSCs) are protective against myocardial injury. The purpose of the study was to investigate the role and mechanisms by which exosomes promote cardiomyocyte survival and function following myocardial infarction (MI). hMSCs were cultured under hypoxic and normoxic conditions. Hypoxiaconditioned hMSC-derived exosomes (Hypo-Exo) and normoxic-conditioned hMSC-derived exosomes (Nor-Exo) were collected and intramyocardially injected into rats with MI. The therapeutic effects of Hypo-Exo and Nor-Exo were evaluated after 4 weeks. Quantitative real-time PCR (qRT-PCR) was used to detect the expression of candidate long noncoding RNA urothelial carcinoma associated 1 (lncRNA-UCA1) in Nor-Exo and Hypo-Exo. Intramyocardial injection of lncRNA-UCA1-knockdown-Hypo-Exo in a rat model of MI was then performed and the cardiac function was characterized. The target and downstream of the molecular mechanism lncRNA-UCA1 was disclosed by luciferase reporter assays and western blot. Circulating exosomal lncRNA-UCA1 level in AMI patients and healthy volunteers was assessed. We found that (1) hMSC exosomal (from hypoxic and normoxic conditions) cardioprotection in vitro and in vivo correlated with the presence of encapsulated lncRNA-UCA1 in exosomes; (2) lncRNA-UCA1 targeted miR-873 via sponging, reducing the latter's suppressive effects on its target XIAP, and this translated into AMPK phosphorylation and increased level of the antiapoptotic protein BCL2; and (3) plasma derived from patients with AMI contained exosomes enriched with the lncRNA-UCA1, unlike that from normal subjects. This study demonstrates that Hypo-Exo lncRNA-UCA1 plays a cardioprotective role via the miR-873-5p/XIAP axis and circulating exosomal lncRNA-UCA1 may be a promising novel biomarker for the diagnosis of AMI.

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Comparison of miR‐16 and cel‐miR‐39 as reference controls for serum miRNA normalization in colorectal cancer

Cited by 15 publications

References 10 publications

Identification of hsa-miR-1275 as a Novel Biomarker Targeting MECP2 for Human Epilepsy of Unknown Etiology

Identification of hsa-miR-1275 as a Novel Biomarker Targeting MECP2 for Human Epilepsy of Unknown Etiology

miRNAs as Biomarkers in Disease: Latest Findings Regarding Their Role in Diagnosis and Prognosis

Long noncoding RNA UCA1 from hypoxia-conditioned hMSC-derived exosomes: a novel molecular target for cardioprotection through miR-873-5p/XIAP axis

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