Cargando…

The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance

BACKGROUND: Autophagy has been found to be involved in the multidrug resistance (MDR) of cancers, but whether it is associated with resistance of small cell lung cancer (SCLC) has not been studied. Here, we hypothesized that a potential autophagy-regulating miRNA, miR-199a-5p, regulated cisplatin-re...

Descripción completa

Detalles Bibliográficos
Autores principales: Li, Tiezhi, Zhang, Helin, Wang, Zhichao, Gao, Shaolin, Zhang, Xu, Zhu, Haiyong, Wang, Na, Li, Honglin
Formato: Online Artículo Texto
Lenguaje:English
Publicado: BioMed Central 2022
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8922820/
https://www.ncbi.nlm.nih.gov/pubmed/35292022
http://dx.doi.org/10.1186/s12935-022-02505-1
_version_ 1784669570908291072
author Li, Tiezhi
Zhang, Helin
Wang, Zhichao
Gao, Shaolin
Zhang, Xu
Zhu, Haiyong
Wang, Na
Li, Honglin
author_facet Li, Tiezhi
Zhang, Helin
Wang, Zhichao
Gao, Shaolin
Zhang, Xu
Zhu, Haiyong
Wang, Na
Li, Honglin
author_sort Li, Tiezhi
collection PubMed
description BACKGROUND: Autophagy has been found to be involved in the multidrug resistance (MDR) of cancers, but whether it is associated with resistance of small cell lung cancer (SCLC) has not been studied. Here, we hypothesized that a potential autophagy-regulating miRNA, miR-199a-5p, regulated cisplatin-resistant SCLC. METHODS: We validated the MDR of H446/EP using CCK-8 and LDH. We tested the binding of miR-199a-5p to p62 using the Dual-Luciferase assay and validated the association of miR-199a-5p and p62 in SCLC samples. We overexpressed (OE) and knocked down (KD) miR-199a-5p in H446 and H446/EP and determined the expression of miR-199a-5p, autophagy-related proteins, and the formation of autophagolysosomes using QPCR, western blotting, and MDC staining respectively. These results were validated in an orthotopic H446 mouse model of SCLC. RESULTS: H446/EP was resistant to cisplatin, etoposide, paclitexal, epirubicin, irinotecan, and vinorelbine. Exposure of cisplatin at 5 μg/ml for 24 h increased LC3II/LC3I, ATG5, p62, and the formation of autophagolysosomes in H446 cells, but not in H446/EP cells. The expression of miR-199a-5p was up-regulated in H446/EP compared to H446. MiR-199a-5p directly targeted the p62 gene. The expression of miR-199a-5p and p62 were correlated in SCLC samples. In H446 and H69PR, the OE of miR-199a-5p increased LC3II/LC3I, p62, and the formation of autophagolysosomes, but not ATG5, while the KD of miR-199a-5p decreased p62, but did not affect LC3II/LC3I, ATG5, and the formation of autophagolysosomes. In H446/EP, the OE of miR-199a-5p decreased p62 only. These results were generally consistent to results in the animal tumor samples. CONCLUSIONS: The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12935-022-02505-1.
format Online
Article
Text
id pubmed-8922820
institution National Center for Biotechnology Information
language English
publishDate 2022
publisher BioMed Central
record_format MEDLINE/PubMed
spelling pubmed-89228202022-03-22 The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance Li, Tiezhi Zhang, Helin Wang, Zhichao Gao, Shaolin Zhang, Xu Zhu, Haiyong Wang, Na Li, Honglin Cancer Cell Int Primary Research BACKGROUND: Autophagy has been found to be involved in the multidrug resistance (MDR) of cancers, but whether it is associated with resistance of small cell lung cancer (SCLC) has not been studied. Here, we hypothesized that a potential autophagy-regulating miRNA, miR-199a-5p, regulated cisplatin-resistant SCLC. METHODS: We validated the MDR of H446/EP using CCK-8 and LDH. We tested the binding of miR-199a-5p to p62 using the Dual-Luciferase assay and validated the association of miR-199a-5p and p62 in SCLC samples. We overexpressed (OE) and knocked down (KD) miR-199a-5p in H446 and H446/EP and determined the expression of miR-199a-5p, autophagy-related proteins, and the formation of autophagolysosomes using QPCR, western blotting, and MDC staining respectively. These results were validated in an orthotopic H446 mouse model of SCLC. RESULTS: H446/EP was resistant to cisplatin, etoposide, paclitexal, epirubicin, irinotecan, and vinorelbine. Exposure of cisplatin at 5 μg/ml for 24 h increased LC3II/LC3I, ATG5, p62, and the formation of autophagolysosomes in H446 cells, but not in H446/EP cells. The expression of miR-199a-5p was up-regulated in H446/EP compared to H446. MiR-199a-5p directly targeted the p62 gene. The expression of miR-199a-5p and p62 were correlated in SCLC samples. In H446 and H69PR, the OE of miR-199a-5p increased LC3II/LC3I, p62, and the formation of autophagolysosomes, but not ATG5, while the KD of miR-199a-5p decreased p62, but did not affect LC3II/LC3I, ATG5, and the formation of autophagolysosomes. In H446/EP, the OE of miR-199a-5p decreased p62 only. These results were generally consistent to results in the animal tumor samples. CONCLUSIONS: The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance. SUPPLEMENTARY INFORMATION: The online version contains supplementary material available at 10.1186/s12935-022-02505-1. BioMed Central 2022-03-15 /pmc/articles/PMC8922820/ /pubmed/35292022 http://dx.doi.org/10.1186/s12935-022-02505-1 Text en © The Author(s) 2022 https://creativecommons.org/licenses/by/4.0/Open AccessThis article is licensed under a Creative Commons Attribution 4.0 International License, which permits use, sharing, adaptation, distribution and reproduction in any medium or format, as long as you give appropriate credit to the original author(s) and the source, provide a link to the Creative Commons licence, and indicate if changes were made. The images or other third party material in this article are included in the article's Creative Commons licence, unless indicated otherwise in a credit line to the material. If material is not included in the article's Creative Commons licence and your intended use is not permitted by statutory regulation or exceeds the permitted use, you will need to obtain permission directly from the copyright holder. To view a copy of this licence, visit http://creativecommons.org/licenses/by/4.0/ (https://creativecommons.org/licenses/by/4.0/) . 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 in a credit line to the data.
spellingShingle Primary Research
Li, Tiezhi
Zhang, Helin
Wang, Zhichao
Gao, Shaolin
Zhang, Xu
Zhu, Haiyong
Wang, Na
Li, Honglin
The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
title The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
title_full The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
title_fullStr The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
title_full_unstemmed The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
title_short The regulation of autophagy by the miR-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
title_sort regulation of autophagy by the mir-199a-5p/p62 axis was a potential mechanism of small cell lung cancer cisplatin resistance
topic Primary Research
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8922820/
https://www.ncbi.nlm.nih.gov/pubmed/35292022
http://dx.doi.org/10.1186/s12935-022-02505-1
work_keys_str_mv AT litiezhi theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT zhanghelin theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT wangzhichao theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT gaoshaolin theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT zhangxu theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT zhuhaiyong theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT wangna theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT lihonglin theregulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT litiezhi regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT zhanghelin regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT wangzhichao regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT gaoshaolin regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT zhangxu regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT zhuhaiyong regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT wangna regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance
AT lihonglin regulationofautophagybythemir199a5pp62axiswasapotentialmechanismofsmallcelllungcancercisplatinresistance