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Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis
BACKGROUND: Recent evidence indicates that metformin inhibits mammalian cancer growth and metastasis through the regulation of microRNAs. Metformin regulates miR-381 stability, which plays a vital role in tumor progression. Moreover, increased YAP expression and activity induce non-small cell lung c...
Autores principales: | , , , , , , , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
Publicado: |
BioMed Central
2020
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Materias: | |
Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6945774/ https://www.ncbi.nlm.nih.gov/pubmed/31906986 http://dx.doi.org/10.1186/s13046-019-1503-6 |
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author | Jin, Dan Guo, Jiwei Wu, Yan Chen, Weiwei Du, Jing Yang, Lijuan Wang, Xiaohong Gong, Kaikai Dai, Juanjuan Miao, Shuang Li, Xuelin Su, Guoming |
author_facet | Jin, Dan Guo, Jiwei Wu, Yan Chen, Weiwei Du, Jing Yang, Lijuan Wang, Xiaohong Gong, Kaikai Dai, Juanjuan Miao, Shuang Li, Xuelin Su, Guoming |
author_sort | Jin, Dan |
collection | PubMed |
description | BACKGROUND: Recent evidence indicates that metformin inhibits mammalian cancer growth and metastasis through the regulation of microRNAs. Metformin regulates miR-381 stability, which plays a vital role in tumor progression. Moreover, increased YAP expression and activity induce non-small cell lung cancer (NSCLC) tumor growth and metastasis. However, the molecular mechanism underpinning how metformin-induced upregulation of miR-381 directly targets YAP or its interactions with the epithelial-mesenchymal transition (EMT) marker protein Snail in NSCLC is still unknown. METHODS: Levels of RNA and protein were analyzed using qPCR, western blotting and immunofluorescence staining. Cellular proliferation was detected using a CCK8 assay. Cell migration and invasion were analyzed using wound healing and transwell assays. Promoter activity and transcription were investigated using the luciferase reporter assay. Chromatin immunoprecipitation was used to detect the binding of YAP to the promoter of Snail. The interaction between miR-381 and the 3′UTR of YAP mRNA was analyzed using the MS2 expression system and co-immunoprecipitation with biotin. RESULTS: We observed that miR-381 expression is negatively correlated with YAP expression and plays an opposite role to YAP in the regulation of cellular proliferation, invasion, migration, and EMT of NSCLC cells. The miR-381 function as a tumor suppressor was significantly downregulated in lung cancer tissue specimens and cell lines, which decreased the expression of its direct target YAP. In addition, metformin decreased cell growth, migration, invasion, and EMT via up-regulation of miR-381. Moreover, YAP, which functions as a co-transcription factor, enhanced NSCLC progression and metastasis by upregulation of Snail. Snail knockdown downregulated the mesenchymal marker vimentin and upregulated the epithelial marker E-cadherin in lung cancer cells. Furthermore, miR-381, YAP, and Snail constitute the miR-381-YAP-Snail signal axis, which is repressed by metformin, and enhances cancer cell invasiveness by directly regulating EMT. CONCLUSIONS: Metformin-induced repression of miR-381-YAP-Snail axis activity disrupts NSCLC growth and metastasis. Thus, we believe that the miR-381-YAP-Snail signal axis may be a suitable diagnostic marker and a potential therapeutic target for lung cancer. |
format | Online Article Text |
id | pubmed-6945774 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2020 |
publisher | BioMed Central |
record_format | MEDLINE/PubMed |
spelling | pubmed-69457742020-01-09 Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis Jin, Dan Guo, Jiwei Wu, Yan Chen, Weiwei Du, Jing Yang, Lijuan Wang, Xiaohong Gong, Kaikai Dai, Juanjuan Miao, Shuang Li, Xuelin Su, Guoming J Exp Clin Cancer Res Research BACKGROUND: Recent evidence indicates that metformin inhibits mammalian cancer growth and metastasis through the regulation of microRNAs. Metformin regulates miR-381 stability, which plays a vital role in tumor progression. Moreover, increased YAP expression and activity induce non-small cell lung cancer (NSCLC) tumor growth and metastasis. However, the molecular mechanism underpinning how metformin-induced upregulation of miR-381 directly targets YAP or its interactions with the epithelial-mesenchymal transition (EMT) marker protein Snail in NSCLC is still unknown. METHODS: Levels of RNA and protein were analyzed using qPCR, western blotting and immunofluorescence staining. Cellular proliferation was detected using a CCK8 assay. Cell migration and invasion were analyzed using wound healing and transwell assays. Promoter activity and transcription were investigated using the luciferase reporter assay. Chromatin immunoprecipitation was used to detect the binding of YAP to the promoter of Snail. The interaction between miR-381 and the 3′UTR of YAP mRNA was analyzed using the MS2 expression system and co-immunoprecipitation with biotin. RESULTS: We observed that miR-381 expression is negatively correlated with YAP expression and plays an opposite role to YAP in the regulation of cellular proliferation, invasion, migration, and EMT of NSCLC cells. The miR-381 function as a tumor suppressor was significantly downregulated in lung cancer tissue specimens and cell lines, which decreased the expression of its direct target YAP. In addition, metformin decreased cell growth, migration, invasion, and EMT via up-regulation of miR-381. Moreover, YAP, which functions as a co-transcription factor, enhanced NSCLC progression and metastasis by upregulation of Snail. Snail knockdown downregulated the mesenchymal marker vimentin and upregulated the epithelial marker E-cadherin in lung cancer cells. Furthermore, miR-381, YAP, and Snail constitute the miR-381-YAP-Snail signal axis, which is repressed by metformin, and enhances cancer cell invasiveness by directly regulating EMT. CONCLUSIONS: Metformin-induced repression of miR-381-YAP-Snail axis activity disrupts NSCLC growth and metastasis. Thus, we believe that the miR-381-YAP-Snail signal axis may be a suitable diagnostic marker and a potential therapeutic target for lung cancer. BioMed Central 2020-01-06 /pmc/articles/PMC6945774/ /pubmed/31906986 http://dx.doi.org/10.1186/s13046-019-1503-6 Text en © The Author(s). 2020 https://creativecommons.org/licenses/by/4.0/Open Access This article is distributed under the terms of the Creative Commons Attribution 4.0 International License (http://creativecommons.org/licenses/by/4.0/ (https://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/ (https://creativecommons.org/publicdomain/zero/1.0/) ) applies to the data made available in this article, unless otherwise stated. |
spellingShingle | Research Jin, Dan Guo, Jiwei Wu, Yan Chen, Weiwei Du, Jing Yang, Lijuan Wang, Xiaohong Gong, Kaikai Dai, Juanjuan Miao, Shuang Li, Xuelin Su, Guoming Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis |
title | Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis |
title_full | Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis |
title_fullStr | Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis |
title_full_unstemmed | Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis |
title_short | Metformin-repressed miR-381-YAP-snail axis activity disrupts NSCLC growth and metastasis |
title_sort | metformin-repressed mir-381-yap-snail axis activity disrupts nsclc growth and metastasis |
topic | Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6945774/ https://www.ncbi.nlm.nih.gov/pubmed/31906986 http://dx.doi.org/10.1186/s13046-019-1503-6 |
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