Cargando…
Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis
PURPOSE: We aimed to identify and verify the key genes and lncRNAs associated with acute lung injury (ALI) and explore the pathogenesis of ALI. Research showed that lower expression of the lncRNA metastasis-associated lung carcinoma transcript 1 (MALAT1) alleviates lung injury induced by lipopolysac...
Autores principales: | , , , , , , , , , |
---|---|
Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
Frontiers Media S.A.
2020
|
Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7575725/ https://www.ncbi.nlm.nih.gov/pubmed/33117815 http://dx.doi.org/10.3389/fcell.2020.586869 |
_version_ | 1783597863804076032 |
---|---|
author | Nan, Chuan-chuan Zhang, Ning Cheung, Kenneth C. P. Zhang, Hua-dong Li, Wei Hong, Cheng-ying Chen, Huai-sheng Liu, Xue-yan Li, Nan Cheng, Lixin |
author_facet | Nan, Chuan-chuan Zhang, Ning Cheung, Kenneth C. P. Zhang, Hua-dong Li, Wei Hong, Cheng-ying Chen, Huai-sheng Liu, Xue-yan Li, Nan Cheng, Lixin |
author_sort | Nan, Chuan-chuan |
collection | PubMed |
description | PURPOSE: We aimed to identify and verify the key genes and lncRNAs associated with acute lung injury (ALI) and explore the pathogenesis of ALI. Research showed that lower expression of the lncRNA metastasis-associated lung carcinoma transcript 1 (MALAT1) alleviates lung injury induced by lipopolysaccharide (LPS). Nevertheless, the mechanisms of MALAT1 on cellular apoptosis remain unclear in LPS-stimulated ALI. We investigated the mechanism of MALAT1 in modulating the apoptosis of LPS-induced human pulmonary alveolar epithelial cells (HPAEpiC). METHODS: Differentially expressed lncRNAs between the ALI samples and normal controls were identified using gene expression profiles. ALI-related genes were determined by the overlap of differentially expressed genes (DEGs), genes correlated with lung, genes correlated with key lncRNAs, and genes sharing significantly high proportions of microRNA targets with MALAT1. Quantitative real-time PCR (qPCR) was applied to detect the expression of MALAT1, microRNA (miR)-194-5p, and forkhead box P2 (FOXP2) mRNA in 1 μg/ml LPS-treated HPAEpiC. MALAT1 knockdown vectors, miR-194-5p inhibitors, and ov-FOXP2 were constructed and used to transfect HPAEpiC. The influence of MALAT1 knockdown on LPS-induced HPAEpiC proliferation and apoptosis via the miR-194-5p/FOXP2 axis was determined using Cell counting kit-8 (CCK-8) assay, flow cytometry, and Western blotting analysis, respectively. The interactions between MALAT1, miR-194-5p, and FOXP2 were verified using dual-luciferase reporter gene assay. RESULTS: We identified a key lncRNA (MALAT1) and three key genes (EYA1, WNT5A, and FOXP2) that are closely correlated with the pathogenesis of ALI. LPS stimulation promoted MALAT1 expression and apoptosis and also inhibited HPAEpiC viability. MALAT1 knockdown significantly improved viability and suppressed the apoptosis of LPS-stimulated HPAEpiC. Moreover, MALAT1 directly targeted miR-194-5p, a downregulated miRNA in LPS-stimulated HPAEpiC, when FOXP2 was overexpressed. MALAT1 knockdown led to the overexpression of miR-194-5p and restrained FOXP2 expression. Furthermore, inhibition of miR-194-5p exerted a rescue effect on MALAT1 knockdown of FOXP2, whereas the overexpression of FOXP2 reversed the effect of MALAT1 knockdown on viability and apoptosis of LPS-stimulated HPAEpiC. CONCLUSION: Our results demonstrated that MALAT1 knockdown alleviated HPAEpiC apoptosis by competitively binding to miR-194-5p and then elevating the inhibitory effect on its target FOXP2. These data provide a novel insight into the role of MALAT1 in the progression of ALI and potential diagnostic and therapeutic strategies for ALI patients. |
format | Online Article Text |
id | pubmed-7575725 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | Frontiers Media S.A. |
record_format | MEDLINE/PubMed |
spelling | pubmed-75757252020-10-27 Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis Nan, Chuan-chuan Zhang, Ning Cheung, Kenneth C. P. Zhang, Hua-dong Li, Wei Hong, Cheng-ying Chen, Huai-sheng Liu, Xue-yan Li, Nan Cheng, Lixin Front Cell Dev Biol Cell and Developmental Biology PURPOSE: We aimed to identify and verify the key genes and lncRNAs associated with acute lung injury (ALI) and explore the pathogenesis of ALI. Research showed that lower expression of the lncRNA metastasis-associated lung carcinoma transcript 1 (MALAT1) alleviates lung injury induced by lipopolysaccharide (LPS). Nevertheless, the mechanisms of MALAT1 on cellular apoptosis remain unclear in LPS-stimulated ALI. We investigated the mechanism of MALAT1 in modulating the apoptosis of LPS-induced human pulmonary alveolar epithelial cells (HPAEpiC). METHODS: Differentially expressed lncRNAs between the ALI samples and normal controls were identified using gene expression profiles. ALI-related genes were determined by the overlap of differentially expressed genes (DEGs), genes correlated with lung, genes correlated with key lncRNAs, and genes sharing significantly high proportions of microRNA targets with MALAT1. Quantitative real-time PCR (qPCR) was applied to detect the expression of MALAT1, microRNA (miR)-194-5p, and forkhead box P2 (FOXP2) mRNA in 1 μg/ml LPS-treated HPAEpiC. MALAT1 knockdown vectors, miR-194-5p inhibitors, and ov-FOXP2 were constructed and used to transfect HPAEpiC. The influence of MALAT1 knockdown on LPS-induced HPAEpiC proliferation and apoptosis via the miR-194-5p/FOXP2 axis was determined using Cell counting kit-8 (CCK-8) assay, flow cytometry, and Western blotting analysis, respectively. The interactions between MALAT1, miR-194-5p, and FOXP2 were verified using dual-luciferase reporter gene assay. RESULTS: We identified a key lncRNA (MALAT1) and three key genes (EYA1, WNT5A, and FOXP2) that are closely correlated with the pathogenesis of ALI. LPS stimulation promoted MALAT1 expression and apoptosis and also inhibited HPAEpiC viability. MALAT1 knockdown significantly improved viability and suppressed the apoptosis of LPS-stimulated HPAEpiC. Moreover, MALAT1 directly targeted miR-194-5p, a downregulated miRNA in LPS-stimulated HPAEpiC, when FOXP2 was overexpressed. MALAT1 knockdown led to the overexpression of miR-194-5p and restrained FOXP2 expression. Furthermore, inhibition of miR-194-5p exerted a rescue effect on MALAT1 knockdown of FOXP2, whereas the overexpression of FOXP2 reversed the effect of MALAT1 knockdown on viability and apoptosis of LPS-stimulated HPAEpiC. CONCLUSION: Our results demonstrated that MALAT1 knockdown alleviated HPAEpiC apoptosis by competitively binding to miR-194-5p and then elevating the inhibitory effect on its target FOXP2. These data provide a novel insight into the role of MALAT1 in the progression of ALI and potential diagnostic and therapeutic strategies for ALI patients. Frontiers Media S.A. 2020-10-07 /pmc/articles/PMC7575725/ /pubmed/33117815 http://dx.doi.org/10.3389/fcell.2020.586869 Text en Copyright © 2020 Nan, Zhang, Cheung, Zhang, Li, Hong, Chen, Liu, Li and Cheng. http://creativecommons.org/licenses/by/4.0/ This is an open-access article distributed under the terms of the Creative Commons Attribution License (CC BY). The use, distribution or reproduction in other forums is permitted, provided the original author(s) and the copyright owner(s) are credited and that the original publication in this journal is cited, in accordance with accepted academic practice. No use, distribution or reproduction is permitted which does not comply with these terms. |
spellingShingle | Cell and Developmental Biology Nan, Chuan-chuan Zhang, Ning Cheung, Kenneth C. P. Zhang, Hua-dong Li, Wei Hong, Cheng-ying Chen, Huai-sheng Liu, Xue-yan Li, Nan Cheng, Lixin Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis |
title | Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis |
title_full | Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis |
title_fullStr | Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis |
title_full_unstemmed | Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis |
title_short | Knockdown of lncRNA MALAT1 Alleviates LPS-Induced Acute Lung Injury via Inhibiting Apoptosis Through the miR-194-5p/FOXP2 Axis |
title_sort | knockdown of lncrna malat1 alleviates lps-induced acute lung injury via inhibiting apoptosis through the mir-194-5p/foxp2 axis |
topic | Cell and Developmental Biology |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7575725/ https://www.ncbi.nlm.nih.gov/pubmed/33117815 http://dx.doi.org/10.3389/fcell.2020.586869 |
work_keys_str_mv | AT nanchuanchuan knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT zhangning knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT cheungkennethcp knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT zhanghuadong knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT liwei knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT hongchengying knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT chenhuaisheng knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT liuxueyan knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT linan knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis AT chenglixin knockdownoflncrnamalat1alleviateslpsinducedacutelunginjuryviainhibitingapoptosisthroughthemir1945pfoxp2axis |