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      Long non-coding maternally expressed gene 3 regulates cigarette smoke extract-induced apoptosis, inflammation and cytotoxicity by sponging miR-181a-2-3p in 16HBE cells

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          Abstract

          Accumulating evidence has suggested that long non-coding (lnc)RNAs are widely involved in the progression of multiple diseases, including chronic obstructive pulmonary disease (COPD). The aim of the present study was to explore the function and molecule mechanism of maternally expressed gene 3 (MEG3) in cigarette smoke extract (CSE)-treated 16HBE cells. Cell viability and apoptosis were evaluated using Cell Counting Kit-8 analysis and flow cytometry, respectively. Western blot analysis was carried out to determine the protein levels of Bcl-2, Bax and cleaved caspase-3. ELISA assays were utilized to measure the protein levels of IL-1β and IL-6 and TNF-α. Cytotoxicity was assessed using a lactate dehydrogenase release assay. The expression levels of MEG3 and microRNA (miR)-181a-2-3p were detected using reverse transcription-quantitative PCR. The interaction between miR-181a-2-3p and MEG3 was predicted using DIANA tools and verified by a dual-luciferase reporter assay and RNA Immunoprecipitation assay. MEG3 expression was enhanced while miR-181a-2-3p abundance was reduced in the serum of patients with COPD and CSE-treated 16HBE cells. MEG3-knockdown or miR-181a-2-3p-overexpression inhibited CSE-induced apoptosis, inflammation and cytotoxicity in 16HBE cells. Moreover, miR-181a-2-3p directly bind to MEG3 and its knockdown reversed the inhibitory effect of MEG3 interference on apoptosis, inflammation and cytotoxicity in CSE-treated 16HBE cells. Overall, MEG3-knockdown suppressed CSE-induced apoptosis, inflammation and cytotoxicity in 16HBE cells by upregulating miR-181a-2-3p, providing a promising therapeutic target for treatment of CSE-induced COPD.

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          Most cited references47

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          Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) Method.

          The two most commonly used methods to analyze data from real-time, quantitative PCR experiments are absolute quantification and relative quantification. Absolute quantification determines the input copy number, usually by relating the PCR signal to a standard curve. Relative quantification relates the PCR signal of the target transcript in a treatment group to that of another sample such as an untreated control. The 2(-Delta Delta C(T)) method is a convenient way to analyze the relative changes in gene expression from real-time quantitative PCR experiments. The purpose of this report is to present the derivation, assumptions, and applications of the 2(-Delta Delta C(T)) method. In addition, we present the derivation and applications of two variations of the 2(-Delta Delta C(T)) method that may be useful in the analysis of real-time, quantitative PCR data. Copyright 2001 Elsevier Science (USA).
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            MicroRNAs: target recognition and regulatory functions.

            MicroRNAs (miRNAs) are endogenous approximately 23 nt RNAs that play important gene-regulatory roles in animals and plants by pairing to the mRNAs of protein-coding genes to direct their posttranscriptional repression. This review outlines the current understanding of miRNA target recognition in animals and discusses the widespread impact of miRNAs on both the expression and evolution of protein-coding genes.
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              A ceRNA hypothesis: the Rosetta Stone of a hidden RNA language?

              Here, we present a unifying hypothesis about how messenger RNAs, transcribed pseudogenes, and long noncoding RNAs "talk" to each other using microRNA response elements (MREs) as letters of a new language. We propose that this "competing endogenous RNA" (ceRNA) activity forms a large-scale regulatory network across the transcriptome, greatly expanding the functional genetic information in the human genome and playing important roles in pathological conditions, such as cancer. Copyright © 2011 Elsevier Inc. All rights reserved.
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                Author and article information

                Journal
                Oncol Lett
                Oncol Lett
                OL
                Oncology Letters
                D.A. Spandidos
                1792-1074
                1792-1082
                January 2021
                17 November 2020
                17 November 2020
                : 21
                : 1
                : 45
                Affiliations
                [1 ]Department of Respiratory Medicine, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China
                [2 ]Department of Pediatrics, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China
                [3 ]Department of Gastroenterology, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China
                [4 ]Department of Critical Care Medicine, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China
                [5 ]Department of Nephrology, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China
                Author notes
                Correspondence to: Dr Shiming Fan, Department of Respiratory Medicine, Changning County Hospital of Traditional Chinese Medicine, 128 Zhuhai Road, Changning, Yibin, Sichuan 644300, P.R. China, E-mail: vzuuti@ 123456163.com
                [*]

                Contributed equally

                Article
                OL-0-0-12306
                10.3892/ol.2020.12306
                7693283
                33262837
                77665b87-7fe7-4cc6-9b60-122e7f8b1924
                Copyright: © Fan et al.

                This is an open access article distributed under the terms of the Creative Commons Attribution-NonCommercial-NoDerivs License, which permits use and distribution in any medium, provided the original work is properly cited, the use is non-commercial and no modifications or adaptations are made.

                History
                : 25 December 2019
                : 26 October 2020
                Categories
                Articles

                Oncology & Radiotherapy
                chronic obstructive pulmonary disease,maternally expressed gene 3,micror-181a-2-3p,cell viability,apoptosis,inflammation,cytotoxicity

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