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      A panel of miRNAs derived from plasma extracellular vesicles as novel diagnostic biomarkers of lung adenocarcinoma

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          Abstract

          Lung cancer is the leading cause of cancer‐related morbidity and mortality worldwide, with lung adenocarcinoma (LUAD) being the most common histological subtype (approximately 40%). In the absence of reliable screening biomarkers for early diagnosis, most patients with LUAD are inevitably diagnosed at an advanced stage. MicroRNAs (miRNAs) encapsulated within plasma‐derived extracellular vesicles (EVs) may be suitable for use as noninvasive diagnostic biomarkers for aggressive malignancies, including LUAD. In this study, we first investigated the miRNA profiles of plasma‐derived EVs from LUAD patients and healthy donors, and then systematically evaluated the expression patterns of selected plasma‐derived EV miRNAs in a large cohort of patients with LUAD and healthy controls. Notably, we observed that miR‐451a, miR‐194‐5p, and miR‐486‐5p were significantly increased in EVs from LUAD patients, compared to healthy controls. The area under the curve values for the three miRNAs were 0.9040 (95% confidence interval [CI], 0.8633–0.9447) for miR‐451a, 0.7492 (95% CI, 0.6992–0.7992) for miR‐194‐5p, and 0.9574 (95% CI, 0.9378–0.9769) for miR‐486‐5p, while the AUC of the combination of these three miRNAs was 0.9650. Thus, these results suggest that these EV miRNAs may be promising candidates for the development of highly effective, noninvasive biomarkers for early LUAD diagnosis.

          Abstract

          Lung cancer is the leading cause of cancer‐related morbidity and mortality worldwide, with lung adenocarcinoma (LUAD) being the most common histological subtype (approximately 40%). In this study, we observed that miR‐451a, miR‐194‐5p, and miR‐486‐5p in extracellular vesicles derived from plasma may be suitable as highly effective and noninvasive biomarkers for early LUAD diagnosis.

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          Most cited references26

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          Haemolysis during Sample Preparation Alters microRNA Content of Plasma

          The presence of cell-free microRNAs (miRNAs) has been detected in a range of body fluids. The miRNA content of plasma/serum in particular has been proposed as a potential source of novel biomarkers for a number of diseases. Nevertheless, the quantification of miRNAs from plasma or serum is made difficult due to inefficient isolation and lack of consensus regarding the optimal reference miRNA. The effect of haemolysis on the quantification and normalisation of miRNAs in plasma has not been investigated in great detail. We found that levels of miR-16, a commonly used reference gene, showed little variation when measured in plasma samples from healthy volunteers or patients with malignant mesothelioma or coronary artery disease. Including samples with evidence of haemolysis led to variation in miR-16 levels and consequently decreased its ability to serve as a reference. The levels of miR-16 and miR-451, both present in significant levels in red blood cells, were proportional to the degree of haemolysis. Measurements of the level of these miRNAs in whole blood, plasma, red blood cells and peripheral blood mononuclear cells revealed that the miRNA content of red blood cells represents the major source of variation in miR-16 and miR-451 levels measured in plasma. Adding lysed red blood cells to non-haemolysed plasma allowed a cut-off level of free haemoglobin to be determined, below which miR-16 and miR-451 levels displayed little variation between individuals. In conclusion, increases in plasma miR-16 and miR-451 are caused by haemolysis. In the absence of haemolysis the levels of both miR-16 and miR-451 are sufficiently constant to serve as normalisers.
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            Extracellular vesicles: masters of intercellular communication and potential clinical interventions.

            Intercellular signaling via extracellular vesicles (EVs) is an underappreciated modality of cell-cell crosstalk that enables cells to convey packages of complex instructions to specific recipient cells. EVs transmit these instructions through their cargoes of multiple proteins, nucleic acids, and specialized lipids, which are derived from their cells of origin and allow for combinatorial effects upon recipient cells. This Review series brings together the recent progress in our understanding of EV signaling in physiological and pathophysiological conditions, highlighting how certain EVs, particularly exosomes, can promote or regulate infections, host immune responses, development, and various diseases - notably cancer. Given the diverse nature of EVs and their abilities to profoundly modulate host cells, this series puts particular emphasis on the clinical applications of EVs as therapeutics and as diagnostic biomarkers.
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              p53-Responsive micrornas 192 and 215 are capable of inducing cell cycle arrest.

              microRNAs provide a novel layer of regulation for gene expression by interfering with the stability and/or translation of specific target mRNAs. Overall levels of microRNAs are frequently down-regulated in cancer cells, and reducing general microRNA processing increases cancerogenesis in transgenic models, suggesting that at least some microRNAs might act as effectors in tumor suppression. Accordingly, the tumor suppressor p53 up-regulates miR-34a, a microRNA that contributes to apoptosis and acute senescence. Here, we used array hybridization to find that p53 induces two additional, mutually related clusters of microRNAs, leading to the up-regulation of miR-192, miR-194, and miR-215. The same microRNAs were detected at high levels in normal colon tissue but were severely reduced in many colon cancer samples. On the other hand, miR-192 and its cousin miR-215 can each contribute to enhanced CDKN1A/p21 levels, colony suppression, cell cycle arrest, and cell detachment from a solid support. These effects were partially dependent on the presence of wild-type p53. Antagonizing endogenous miR-192 attenuated 5-fluorouracil-induced accumulation of p21. Hence, miR-192 and miR-215 can act as effectors as well as regulators of p53; they seem to suppress cancerogenesis through p21 accumulation and cell cycle arrest.
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                Author and article information

                Contributors
                liuming_nju@163.com
                qzhao@nju.edu.cn
                Journal
                FEBS Open Bio
                FEBS Open Bio
                10.1002/(ISSN)2211-5463
                FEB4
                FEBS Open Bio
                John Wiley and Sons Inc. (Hoboken )
                2211-5463
                21 November 2019
                December 2019
                : 9
                : 12 ( doiID: 10.1002/feb4.v9.12 )
                : 2149-2158
                Affiliations
                [ 1 ] The State Key Laboratory of Pharmaceutical Biotechnology School of Life Sciences Nanjing University China
                [ 2 ] Department of Oncology The First Affiliated Hospital of Nanjing Medical University China
                Author notes
                [*] [* ] Correspondence

                Q. Zhao or M. Liu, State Key Laboratory of Pharmaceutical Biotechnology, School of Life Sciences, Nanjing University, 163 Xianlin Avenue, Nanjing, 210023, China

                Tel/Fax: 86‐25‐89687251

                E‐mails: qzhao@ 123456nju.edu.cn or liuming_nju@ 123456163.com

                Author information
                https://orcid.org/0000-0002-6657-2760
                Article
                FEB412753
                10.1002/2211-5463.12753
                6886307
                31677346
                28001eb5-1278-4f75-95d3-31e4f33adb73
                © 2019 The Authors. Published by FEBS Press and John Wiley & Sons Ltd.

                This is an open access article under the terms of the http://creativecommons.org/licenses/by/4.0/ License, which permits use, distribution and reproduction in any medium, provided the original work is properly cited.

                History
                : 21 September 2019
                : 31 October 2019
                Page count
                Figures: 4, Tables: 2, Pages: 10, Words: 6473
                Funding
                Funded by: National Natural Science Foundation of China
                Award ID: 31701191
                Award ID: 31770809
                Categories
                Research Article
                Research Articles
                Custom metadata
                2.0
                feb412753
                December 2019
                Converter:WILEY_ML3GV2_TO_NLMPMC version:5.7.2 mode:remove_FC converted:02.12.2019

                biomarker,extracellular vesicles,lung adenocarcinoma,mirnas

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