Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer

BACKGROUND: Circular RNAs (circRNAs) are non-coding RNAs with a circular structure that have recently emerged as important regulators of tumorogenesis. Recently, several circRNAS, including circ-10720 have been related to epithelial-mesenchymal transition (EMT) process. In the present study, we have...

Descripción completa

Detalles Bibliográficos
Autores principales: Martín, Jara, Castellano, Joan Josep, Marrades, Ramón María, Canals, Jordi, Viñolas, Nuria, Díaz, Tania, Molins, Laureano, Martinez, Daniel, Han, Bing, Moisés, Jorge, He, Yangyi, Monzó, Mariano, Navarro, Alfons
Formato: Online Artículo Texto
Lenguaje:English
Publicado: AME Publishing Company 2021
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8107756/
https://www.ncbi.nlm.nih.gov/pubmed/34012794
http://dx.doi.org/10.21037/tlcr-20-920
_version_ 1783690007238672384
author Martín, Jara
Castellano, Joan Josep
Marrades, Ramón María
Canals, Jordi
Viñolas, Nuria
Díaz, Tania
Molins, Laureano
Martinez, Daniel
Han, Bing
Moisés, Jorge
He, Yangyi
Monzó, Mariano
Navarro, Alfons
author_facet Martín, Jara
Castellano, Joan Josep
Marrades, Ramón María
Canals, Jordi
Viñolas, Nuria
Díaz, Tania
Molins, Laureano
Martinez, Daniel
Han, Bing
Moisés, Jorge
He, Yangyi
Monzó, Mariano
Navarro, Alfons
author_sort Martín, Jara
collection PubMed
description BACKGROUND: Circular RNAs (circRNAs) are non-coding RNAs with a circular structure that have recently emerged as important regulators of tumorogenesis. Recently, several circRNAS, including circ-10720 have been related to epithelial-mesenchymal transition (EMT) process. In the present study, we have analyzed the role of circ-10720 in non-small-cell lung cancer (NSCLC) and studied its prognostic relevance in resected stage I–IIIa NSCLC patients. METHODS: Circ-10720 expression was analyzed using a custom TaqMan assay in four NSCLC cell lines (HCC44, A549, H23 and H1299) and in the normal immortalized lung cell line BEAS2B. Silencing of circ-10720 was performed using two custom siRNAs which were transfected using lipofectamine 2000. Protein levels were evaluated by Western blot and immunofluorescence. Wound healing and invasion assays were performed to evaluate the impact the circRNA on cell motility. Apoptosis was analyzed by evaluation of Caspase 3–7 activity and proliferation by MTS assay. Moreover, the expression levels of the circRNA were studied in 119 resected NSCLC patients. The expression in tumor tissue was correlated with the main clinicopathological characteristics and with time to relapse (TTR). RESULTS: Circ-10720 was overexpressed in HCC44 and A549 and underexpressed in H23 and H1299 NSCLC cell lines in comparison to BEAS2B normal immortalized lung cell line. CircRNA knockdown in the two circ-10720 overexpressing cell lines was associated with a decrease of Vimentin (VIM) and an increase of E-cadherin (CDH1) protein levels, loss of mesenchymal phenotype, and a significant reduction of migration and invasion capacity. After silencing circ-10720, the apoptosis rate increased and the proliferation was significantly reduced. Furthermore, circ-10720 was upregulated in tumor vs. normal tissue from 119 resected NSCLC patients. In the group of patients not receiving adjuvant treatment, those with high levels of circ-10720 had a shorter TTR than those with low levels and emerged as an independent prognostic value in the multivariate analysis. In tumor tissue, circ-10720 levels positively correlated with the EMT gene Twist1 levels. CONCLUSIONS: Circ-10720 regulates EMT, apoptosis and proliferation and acts as a biomarker of relapse in NSCLC.
format Online
Article
Text
id pubmed-8107756
institution National Center for Biotechnology Information
language English
publishDate 2021
publisher AME Publishing Company
record_format MEDLINE/PubMed
spelling pubmed-81077562021-05-18 Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer Martín, Jara Castellano, Joan Josep Marrades, Ramón María Canals, Jordi Viñolas, Nuria Díaz, Tania Molins, Laureano Martinez, Daniel Han, Bing Moisés, Jorge He, Yangyi Monzó, Mariano Navarro, Alfons Transl Lung Cancer Res Original Article BACKGROUND: Circular RNAs (circRNAs) are non-coding RNAs with a circular structure that have recently emerged as important regulators of tumorogenesis. Recently, several circRNAS, including circ-10720 have been related to epithelial-mesenchymal transition (EMT) process. In the present study, we have analyzed the role of circ-10720 in non-small-cell lung cancer (NSCLC) and studied its prognostic relevance in resected stage I–IIIa NSCLC patients. METHODS: Circ-10720 expression was analyzed using a custom TaqMan assay in four NSCLC cell lines (HCC44, A549, H23 and H1299) and in the normal immortalized lung cell line BEAS2B. Silencing of circ-10720 was performed using two custom siRNAs which were transfected using lipofectamine 2000. Protein levels were evaluated by Western blot and immunofluorescence. Wound healing and invasion assays were performed to evaluate the impact the circRNA on cell motility. Apoptosis was analyzed by evaluation of Caspase 3–7 activity and proliferation by MTS assay. Moreover, the expression levels of the circRNA were studied in 119 resected NSCLC patients. The expression in tumor tissue was correlated with the main clinicopathological characteristics and with time to relapse (TTR). RESULTS: Circ-10720 was overexpressed in HCC44 and A549 and underexpressed in H23 and H1299 NSCLC cell lines in comparison to BEAS2B normal immortalized lung cell line. CircRNA knockdown in the two circ-10720 overexpressing cell lines was associated with a decrease of Vimentin (VIM) and an increase of E-cadherin (CDH1) protein levels, loss of mesenchymal phenotype, and a significant reduction of migration and invasion capacity. After silencing circ-10720, the apoptosis rate increased and the proliferation was significantly reduced. Furthermore, circ-10720 was upregulated in tumor vs. normal tissue from 119 resected NSCLC patients. In the group of patients not receiving adjuvant treatment, those with high levels of circ-10720 had a shorter TTR than those with low levels and emerged as an independent prognostic value in the multivariate analysis. In tumor tissue, circ-10720 levels positively correlated with the EMT gene Twist1 levels. CONCLUSIONS: Circ-10720 regulates EMT, apoptosis and proliferation and acts as a biomarker of relapse in NSCLC. AME Publishing Company 2021-04 /pmc/articles/PMC8107756/ /pubmed/34012794 http://dx.doi.org/10.21037/tlcr-20-920 Text en 2021 Translational Lung Cancer Research. All rights reserved. https://creativecommons.org/licenses/by-nc-nd/4.0/Open Access Statement: This is an Open Access article distributed in accordance with the Creative Commons Attribution-NonCommercial-NoDerivs 4.0 International License (CC BY-NC-ND 4.0), which permits the non-commercial replication and distribution of the article with the strict proviso that no changes or edits are made and the original work is properly cited (including links to both the formal publication through the relevant DOI and the license). See: https://creativecommons.org/licenses/by-nc-nd/4.0 (https://creativecommons.org/licenses/by-nc-nd/4.0/) .
spellingShingle Original Article
Martín, Jara
Castellano, Joan Josep
Marrades, Ramón María
Canals, Jordi
Viñolas, Nuria
Díaz, Tania
Molins, Laureano
Martinez, Daniel
Han, Bing
Moisés, Jorge
He, Yangyi
Monzó, Mariano
Navarro, Alfons
Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer
title Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer
title_full Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer
title_fullStr Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer
title_full_unstemmed Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer
title_short Role of the epithelial-mesenchymal transition-related circular RNA, circ-10720, in non-small-cell lung cancer
title_sort role of the epithelial-mesenchymal transition-related circular rna, circ-10720, in non-small-cell lung cancer
topic Original Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC8107756/
https://www.ncbi.nlm.nih.gov/pubmed/34012794
http://dx.doi.org/10.21037/tlcr-20-920
work_keys_str_mv AT martinjara roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT castellanojoanjosep roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT marradesramonmaria roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT canalsjordi roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT vinolasnuria roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT diaztania roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT molinslaureano roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT martinezdaniel roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT hanbing roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT moisesjorge roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT heyangyi roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT monzomariano roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer
AT navarroalfons roleoftheepithelialmesenchymaltransitionrelatedcircularrnacirc10720innonsmallcelllungcancer