Serum starvation induces cell death in NSCLC via miR-224
Purpose: Increasing evidence suggests that microRNAs (miRNAs) may be involved in the occurrence and progression of non-small cell lung cancer (NSCLC). In the present study, we used serum-starved A549 cells emulating tumor under a nutrient depletion stress in the microenvironment. Patients and method...
Autores principales: | , , , , , |
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Formato: | Online Artículo Texto |
Lenguaje: | English |
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Dove
2019
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Acceso en línea: | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6535431/ https://www.ncbi.nlm.nih.gov/pubmed/31190892 http://dx.doi.org/10.2147/OTT.S186613 |
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author | Wang, Guoqin Han, Jiangqiong Zhuang, Li Li, Shijuan Gong, Quan Chen, Yunlan |
author_facet | Wang, Guoqin Han, Jiangqiong Zhuang, Li Li, Shijuan Gong, Quan Chen, Yunlan |
author_sort | Wang, Guoqin |
collection | PubMed |
description | Purpose: Increasing evidence suggests that microRNAs (miRNAs) may be involved in the occurrence and progression of non-small cell lung cancer (NSCLC). In the present study, we used serum-starved A549 cells emulating tumor under a nutrient depletion stress in the microenvironment. Patients and methods: We first detected the expression level of miR-224 between tumor tissues and the adjacent normal tissues. We analyzed the expression levels of miR-224 and its predicted target phosphatase and tensin homolog (PTEN) using quantitative real-time PCR (qRT-PCR) in starved A549 cells. Following transfection with miR-224 mimic or inhibitor in starved A549 cells, MTT assay, Annexin V FITC/PI staining, and LC-3 immunofluorence staining were performed to investigate the roles of miR-224 on proliferation, apoptosis, and autophagy. Next, the expression of apoptosis-related protein Bax and Bcl-2, autophagy-related proteins LC3, PI3K signaling, and target PTEN were measured using qRT-PCR and Western blot assays. The direct interaction between miR-224 and PTEN was validated with a dual luciferase assay. Results: We found that the expression level of miR-224 in tumor tissues was significantly higher when compared with the adjacent normal tissues. We discovered a reciprocal expression pattern between miR-224 and PTEN in starved A549 cells, and transfection with miR-224 mimic led to down-regulation of PTEN. A dual luciferase assay further confirmed the direct interaction between miR-224 and 3ʹUTR of PTEN. Transfection with miR-224 mimic in starved A549 cells resulted in enhanced cell proliferation, reduced apoptosis, and autophagy, accompanied by increased expression of anti-apoptotic protein Bcl-2, decreased expression of pro-apoptotic protein Bax, and autophagy-related protein LC3. Activation of PI3K was observed in miR-224 mimic transfected cells. The reverse effects by the miR-224 inhibitor in all experiments were observed. Conclusion: Taken together, we proved that miR-224 might play essential roles in cellular functions of nutrient-depleted A549 cells possibly through regulating the target PTEN and downstream signal PI3K, suggesting the potential of miR-224 to be a therapeutic target for NSCLC therapy. |
format | Online Article Text |
id | pubmed-6535431 |
institution | National Center for Biotechnology Information |
language | English |
publishDate | 2019 |
publisher | Dove |
record_format | MEDLINE/PubMed |
spelling | pubmed-65354312019-06-12 Serum starvation induces cell death in NSCLC via miR-224 Wang, Guoqin Han, Jiangqiong Zhuang, Li Li, Shijuan Gong, Quan Chen, Yunlan Onco Targets Ther Original Research Purpose: Increasing evidence suggests that microRNAs (miRNAs) may be involved in the occurrence and progression of non-small cell lung cancer (NSCLC). In the present study, we used serum-starved A549 cells emulating tumor under a nutrient depletion stress in the microenvironment. Patients and methods: We first detected the expression level of miR-224 between tumor tissues and the adjacent normal tissues. We analyzed the expression levels of miR-224 and its predicted target phosphatase and tensin homolog (PTEN) using quantitative real-time PCR (qRT-PCR) in starved A549 cells. Following transfection with miR-224 mimic or inhibitor in starved A549 cells, MTT assay, Annexin V FITC/PI staining, and LC-3 immunofluorence staining were performed to investigate the roles of miR-224 on proliferation, apoptosis, and autophagy. Next, the expression of apoptosis-related protein Bax and Bcl-2, autophagy-related proteins LC3, PI3K signaling, and target PTEN were measured using qRT-PCR and Western blot assays. The direct interaction between miR-224 and PTEN was validated with a dual luciferase assay. Results: We found that the expression level of miR-224 in tumor tissues was significantly higher when compared with the adjacent normal tissues. We discovered a reciprocal expression pattern between miR-224 and PTEN in starved A549 cells, and transfection with miR-224 mimic led to down-regulation of PTEN. A dual luciferase assay further confirmed the direct interaction between miR-224 and 3ʹUTR of PTEN. Transfection with miR-224 mimic in starved A549 cells resulted in enhanced cell proliferation, reduced apoptosis, and autophagy, accompanied by increased expression of anti-apoptotic protein Bcl-2, decreased expression of pro-apoptotic protein Bax, and autophagy-related protein LC3. Activation of PI3K was observed in miR-224 mimic transfected cells. The reverse effects by the miR-224 inhibitor in all experiments were observed. Conclusion: Taken together, we proved that miR-224 might play essential roles in cellular functions of nutrient-depleted A549 cells possibly through regulating the target PTEN and downstream signal PI3K, suggesting the potential of miR-224 to be a therapeutic target for NSCLC therapy. Dove 2019-05-22 /pmc/articles/PMC6535431/ /pubmed/31190892 http://dx.doi.org/10.2147/OTT.S186613 Text en © 2019 Wang et al. http://creativecommons.org/licenses/by-nc/3.0/ This work is published and licensed by Dove Medical Press Limited. The full terms of this license are available at https://www.dovepress.com/terms.php and incorporate the Creative Commons Attribution – Non Commercial (unported, v3.0) License (http://creativecommons.org/licenses/by-nc/3.0/). By accessing the work you hereby accept the Terms. Non-commercial uses of the work are permitted without any further permission from Dove Medical Press Limited, provided the work is properly attributed. For permission for commercial use of this work, please see paragraphs 4.2 and 5 of our Terms (https://www.dovepress.com/terms.php). |
spellingShingle | Original Research Wang, Guoqin Han, Jiangqiong Zhuang, Li Li, Shijuan Gong, Quan Chen, Yunlan Serum starvation induces cell death in NSCLC via miR-224 |
title | Serum starvation induces cell death in NSCLC via miR-224 |
title_full | Serum starvation induces cell death in NSCLC via miR-224 |
title_fullStr | Serum starvation induces cell death in NSCLC via miR-224 |
title_full_unstemmed | Serum starvation induces cell death in NSCLC via miR-224 |
title_short | Serum starvation induces cell death in NSCLC via miR-224 |
title_sort | serum starvation induces cell death in nsclc via mir-224 |
topic | Original Research |
url | https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6535431/ https://www.ncbi.nlm.nih.gov/pubmed/31190892 http://dx.doi.org/10.2147/OTT.S186613 |
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