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      Genome-wide study of salivary microRNAs as potential noninvasive biomarkers for detection of nasopharyngeal carcinoma

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          Abstract

          Background

          Recent studies reported that blood-based microRNAs (miRNAs) could detect cancers and predict prognosis have opened a new field of utilizing circulating miRNAs as cancer biomarkers. In this pilot study, we conducted for the first time, to our knowledge, the evaluation of the applicability of salivary miRNAs as novel biomarkers for nasopharyngeal carcinoma (NPC) detection.

          Methods

          Microarray miRNA expression profiling was performed on saliva samples from 22 newly diagnosed NPC patients and 25 healthy controls, and 12 significantly down-regulated miRNAs were selected for quantitative real-time-PCR (qRT-PCR) validation and further analysis. Their target genes enriched by gene ontology and pathway analysis were used to construct regulatory and interaction networks. The receiver operating characteristic analyses (ROC) and logistic regression were calculated to assess discriminatory accuracy.

          Results

          Twelve dysregulated miRNAs screened by microarray that showed the same expression patterns with qRT-PCR analysis. Through bioinformatics analysis, the most prominent hub gene probably regulated by the 12 down-regulated miRNAs is found to be TP53. The ROC including the 12 miRNAs separated NPC patients from healthy controls with very high accuracy (areas under the receiver operating characteristic curve [AUC] = 0.999, sensitivity = 100.00%, specificity = 96.00%). Furthermore, if only six significantly dysregulated miRNAs were selected for the ROC analysis, the accuracy is still impressive (AUC = 0.941, sensitivity = 95.45%, specificity = 80.00%).

          Conclusions

          This study highlights the potential for salivary miRNAs as biomarkers for the detection of NPC. Meanwhile, differentially expressed miRNAs in saliva might play critical roles in NPC by regulating their target genes, which associated with some significant pathways, such as p53 signaling pathway.

          Electronic supplementary material

          The online version of this article (10.1186/s12885-019-6037-y) contains supplementary material, which is available to authorized users.

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          Most cited references17

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          MicroRNAs in development and disease.

          MicroRNAs (miRNAs) are a class of posttranscriptional regulators that have recently introduced an additional level of intricacy to our understanding of gene regulation. There are currently over 10,000 miRNAs that have been identified in a range of species including metazoa, mycetozoa, viridiplantae, and viruses, of which 940, to date, are found in humans. It is estimated that more than 60% of human protein-coding genes harbor miRNA target sites in their 3' untranslated region and, thus, are potentially regulated by these molecules in health and disease. This review will first briefly describe the discovery, structure, and mode of function of miRNAs in mammalian cells, before elaborating on their roles and significance during development and pathogenesis in the various mammalian organs, while attempting to reconcile their functions with our existing knowledge of their targets. Finally, we will summarize some of the advances made in utilizing miRNAs in therapeutics.
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            Proposal for the 8th edition of the AJCC/UICC staging system for nasopharyngeal cancer in the era of intensity-modulated radiotherapy.

            An accurate staging system is crucial for cancer management. Evaluations for continual suitability and improvement are needed as staging and treatment methods evolve.
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              MicroRNA 29c is down-regulated in nasopharyngeal carcinomas, up-regulating mRNAs encoding extracellular matrix proteins.

              Using highly sensitive microarray-based procedures, we identified eight microRNAs (miRNAs) showing robust differential expression between 31 laser-capture-microdissected nasopharyngeal carcinomas (NPCs) and 10 normal healthy nasopharyngeal epithelial samples. In particular, miRNA mir-29c was expressed at one-fifth the levels in tumors as in normal epithelium. In NPC tumors, the lower mir-29c levels correlated with higher levels of multiple mRNAs whose 3' UTRs can bind mir-29c at target sequences conserved across many vertebrates. In cultured cells, introduction of mir-29c down-regulated these genes at the level of mRNA and inhibited expression of luciferase encoded by vectors having the 3' UTRs of these genes. Moreover, for each of several genes tested, mutating the mir-29c target sites in the 3' UTR abrogated mir-29c-induced inhibition of luciferase expression. Most of the mir-29c-targeted genes identified encode extracellular matrix proteins, including multiple collagens and laminin gamma1, that are associated with tumor cell invasiveness and metastatic potential, prominent characteristics of NPC. Thus, we identify eight miRNAs differentially expressed in NPC and demonstrate the involvement of one in regulating genes involved in metastasis.
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                Author and article information

                Contributors
                wulirong126@126.com
                kxzheng2007@sinano.ac.cn
                cyan2017@sinano.ac.cn
                Panxuan0214@163.com
                105674387@qq.com
                2403083217@qq.com
                wangfeijiang1964@126.com
                gwj006499@sina.com
                hexia206@yeah.net
                jli2006@sinano.ac.cn
                yshen2010@sinano.ac.cn
                Journal
                BMC Cancer
                BMC Cancer
                BMC Cancer
                BioMed Central (London )
                1471-2407
                28 August 2019
                28 August 2019
                2019
                : 19
                : 843
                Affiliations
                [1 ]ISNI 0000 0004 1764 4566, GRID grid.452509.f, Department of Radiation Oncology, , Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & The Affiliated Cancer Hospital of Nanjing Medical University, ; Nanjing, 210009 China
                [2 ]ISNI 0000 0004 1806 6323, GRID grid.458499.d, Nano-Bio-Chem Centre, , Suzhou Institute of Nano-Tech and Nano-Bionics, Chinese Academy of Sciences, ; Suzhou, 215123 China
                [3 ]ISNI 0000 0004 1764 4566, GRID grid.452509.f, Department of Oncology, , Jiangsu Cancer Hospital & Jiangsu Institute of Cancer Research & The Affiliated Cancer Hospital of Nanjing Medical University, ; Nanjing, 210009 China
                Author information
                http://orcid.org/0000-0002-4448-2158
                Article
                6037
                10.1186/s12885-019-6037-y
                6712819
                31455274
                b43fae83-cd52-4277-84a2-c15b2a3c0a6a
                © The Author(s). 2019

                Open AccessThis article is distributed under the terms of the Creative Commons Attribution 4.0 International License ( http://creativecommons.org/licenses/by/4.0/), which permits unrestricted use, distribution, and reproduction in any medium, provided you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons license, and indicate if changes were made. The Creative Commons Public Domain Dedication waiver ( http://creativecommons.org/publicdomain/zero/1.0/) applies to the data made available in this article, unless otherwise stated.

                History
                : 24 March 2019
                : 13 August 2019
                Funding
                Funded by: FundRef http://dx.doi.org/10.13039/501100001809, National Natural Science Foundation of China;
                Award ID: 61671445
                Award ID: 81672989
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100010909, Young Scientists Fund;
                Award ID: 31300301
                Award Recipient :
                Funded by: Jiangsu Provincial Commission of Health and Family Planning (CN)
                Award ID: Q201601
                Award Recipient :
                Funded by: FundRef http://dx.doi.org/10.13039/501100004610, Science and Technology Support Program of Jiangsu Province;
                Award ID: BE2016681
                Award Recipient :
                Categories
                Research Article
                Custom metadata
                © The Author(s) 2019

                Oncology & Radiotherapy
                nasopharyngeal carcinoma,biomarkers,microrna,saliva
                Oncology & Radiotherapy
                nasopharyngeal carcinoma, biomarkers, microrna, saliva

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