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      Expression of Serum Exosomal and Esophageal MicroRNA in Rat Reflux Esophagitis

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          Abstract

          Gastroesophageal reflux disease (GERD) is a common upper gastrointestinal disease. However, the role of exosomal microRNAs (miRNAs) and esophageal miRNAs in GERD has not been studied. A rat model of acid reflux esophagitis was used to establish a novel diagnosis marker for GERD and examine dynamics of miRNA expression in GERD. Rats were sacrificed 3 (acute phase), 7 (sub-acute phase) and 21 days (chronic phase) after induction of esophagitis. Exosomes were extracted from serum, and the expression patterns of serum miRNAs were analyzed. Four upregulated miRNAs (miR-29a-3p, 128-3p, 223-3p and 3473) were identified by microarray analysis. The expression levels of exosomal miR-29a-3p were significantly higher in the chronic phase of reflux esophagitis compared with controls, and increased expression of miR-29a-3p was specific to chronic reflux esophagitis. Esophageal miR-223-3p expression was higher compared with controls, and gradually decreased from acute to chronic phase in esophagitis. In conclusion, exosomal miR-29a-3p and esophageal miR-223-3p might play roles in GERD.

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          Most cited references22

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          Membrane-derived microvesicles: important and underappreciated mediators of cell-to-cell communication.

          Normal and malignant cells shed from their surface membranes as well as secrete from the endosomal membrane compartment circular membrane fragments called microvesicles (MV). MV that are released from viable cells are usually smaller in size compared to the apoptotic bodies derived from damaged cells and unlike them do not contain fragmented DNA. Growing experimental evidence indicates that MV are an underappreciated component of the cell environment and play an important pleiotropic role in many biological processes. Generally, MV are enriched in various bioactive molecules and may (i) directly stimulate cells as a kind of 'signaling complex', (ii) transfer membrane receptors, proteins, mRNA and organelles (e.g., mitochondria) between cells and finally (iii) deliver infectious agents into cells (e.g., human immuno deficiency virus, prions). In this review, we discuss the pleiotropic effects of MV that are important for communication between cells, as well as the role of MV in carcinogenesis, coagulation, immune responses and modulation of susceptibility/infectability of cells to retroviruses or prions.
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            Phylogenetic shadowing and computational identification of human microRNA genes.

            We sequenced 122 miRNAs in 10 primate species to reveal conservation characteristics of miRNA genes. Strong conservation is observed in stems of miRNA hairpins and increased variation in loop sequences. Interestingly, a striking drop in conservation was found for sequences immediately flanking the miRNA hairpins. This characteristic profile was employed to predict novel miRNAs using cross-species comparisons. Nine hundred and seventy-six candidate miRNAs were identified by scanning whole-genome human/mouse and human/rat alignments. Most of the novel candidates are conserved also in other vertebrates (dog, cow, chicken, opossum, zebrafish). Northern blot analysis confirmed the expression of mature miRNAs for 16 out of 69 representative candidates. Additional support for the expression of 179 novel candidates can be found in public databases, their presence in gene clusters, and literature that appeared after these predictions were made. Taken together, these results suggest the presence of significantly higher numbers of miRNAs in the human genome than previously estimated.
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              Clinical impact of serum exosomal microRNA-21 as a clinical biomarker in human esophageal squamous cell carcinoma.

              Exosomes are 40-nm to 100-nm membrane vesicles that are secreted by various cells, and they play a major role in cell-cell communication. The objective of this study was to clarify the significance of the levels of microRNA in exosomes extracted from the sera of patients with esophageal squamous cell cancer (ESCC). The authors isolated exosomes in serum samples from patients who had ESCC and from patients who had benign diseases without systemic inflammation. Total RNA was purified from the exosomes, and expression levels of microRNA-21 (miR-21) were analyzed by quantitative real-time polymerase chain reaction. Serum exosomes from patients with ESCC induced the proliferation of ESCC cells in vitro. The expression levels of exosomal miR-21 were significantly higher in patients with ESCC than those with benign diseases with and without (C-reactive protein <0.3 mg/dL) systemic inflammation. MiR-21 was not detected in serum that remained after exosome extraction. Exosomal miR-21 expression was correlated with advanced tumor classification, positive lymph node status, and the presence of metastasis with inflammation or and clinical stage without inflammation (C-reactive protein <0.3 mg/dL). The current results confirmed that exosomal miR-21 expression is up-regulated in serum from patients with ESCC versus serum from patients who have benign diseases without systemic inflammation. Exosomal miR-21 was positively correlated with tumor progression and aggressiveness, suggesting that it may be a useful target for cancer therapy. Cancer 2013. © 2012 American Cancer Society. Copyright © 2012 American Cancer Society.
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                Author and article information

                Journal
                Int J Mol Sci
                Int J Mol Sci
                ijms
                International Journal of Molecular Sciences
                MDPI
                1422-0067
                25 July 2017
                August 2017
                : 18
                : 8
                : 1611
                Affiliations
                [1 ]Department of Gastroenterology, Osaka City University Graduate School of Medicine, Osaka 545-8585, Japan; uemurarisa@ 123456med.osaka-cu.ac.jp (R.U.); m2075060@ 123456med.osaka-cu.ac.jp (A.H.); m1164972@ 123456med.osaka-cu.ac.jp (A.S.); kojiotani@ 123456med.osaka-cu.ac.jp (K.O.); koichit0802@ 123456gmail.com (K.T.); m1265271@ 123456med.osaka-cu.ac.jp (S.H.); yasuaki1975@ 123456gmail.com (Y.N.); m2079981@ 123456med.osaka-cu.ac.jp (F.T.); nkamata@ 123456med.osaka-cu.ac.jp (N.K.); yamagami@ 123456med.osaka-cu.ac.jp (H.Y.); ttanigawa@ 123456med.osaka-cu.ac.jp (T.T.); watanabet@ 123456med.osaka-cu.ac.jp (T.W.)
                [2 ]Department of Hepatology, Osaka City University Graduate School of Medicine, Osaka 545-8585, Japan; yoshimurak@ 123456med.osaka-cu.ac.jp
                [3 ]Department of Physics, Chuo University, Tokyo 112-8551, Japan; tag@ 123456granular.com
                Author notes
                [* ]Correspondence: yasu@ 123456med.osaka-cu.ac.jp ; Tel.: +81-6-6645-3810
                Author information
                https://orcid.org/0000-0003-0867-8986
                Article
                ijms-18-01611
                10.3390/ijms18081611
                5578003
                28757556
                ddce40e7-c0b9-416d-a7e9-a81a86490f7e
                © 2017 by the authors.

                Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license ( http://creativecommons.org/licenses/by/4.0/).

                History
                : 09 June 2017
                : 20 July 2017
                Categories
                Article

                Molecular biology
                gastroesophageal reflux disease,non-erosive reflux disease,reflux esophagitis,microrna,exosome

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