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

Fan,Shiming; Ren,Yan; Zhang,Wenli; Zhang,Huawei; Wang,Cheng

doi:10.3892/ol.2020.12306

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

Authors:
- Shiming Fan
- Yan Ren
- Wenli Zhang
- Huawei Zhang
- Cheng Wang
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Affiliations: Department of Respiratory Medicine, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China, Department of Pediatrics, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China, Department of Gastroenterology, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China, Department of Critical Care Medicine, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China, Department of Nephrology, Changning County Hospital of Traditional Chinese Medicine, Yibin, Sichuan 644300, P.R. China
Published online on: November 17, 2020 https://doi.org/10.3892/ol.2020.12306
Article Number: 45
Copyright: © Fan et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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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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