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LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk

Although papillary thyroid carcinoma (PTC) has a good prognosis, 20–90% of patients show metastasis to regional lymph nodes and 10–15% of patients show metastasis to distant sites. Metastatic disease represents the main clinical challenge that impacts survival rate. We previously showed that LIMD2 w...

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Autores principales: Araldi, Rodrigo Pinheiro, de Melo, Thatiana Correa, Levy, Débora, de Souza, Dener Madeiro, Maurício, Beatriz, Colozza-Gama, Gabriel Avelar, Bydlowski, Sergio Paulo, Peng, Hongzhuang, Rauscher, Frank J., Cerutti, Janete Maria
Formato: Online Artículo Texto
Lenguaje:English
Publicado: MDPI 2020
Materias:
Acceso en línea:https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7700534/
https://www.ncbi.nlm.nih.gov/pubmed/33238381
http://dx.doi.org/10.3390/cells9112522
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author Araldi, Rodrigo Pinheiro
de Melo, Thatiana Correa
Levy, Débora
de Souza, Dener Madeiro
Maurício, Beatriz
Colozza-Gama, Gabriel Avelar
Bydlowski, Sergio Paulo
Peng, Hongzhuang
Rauscher, Frank J.
Cerutti, Janete Maria
author_facet Araldi, Rodrigo Pinheiro
de Melo, Thatiana Correa
Levy, Débora
de Souza, Dener Madeiro
Maurício, Beatriz
Colozza-Gama, Gabriel Avelar
Bydlowski, Sergio Paulo
Peng, Hongzhuang
Rauscher, Frank J.
Cerutti, Janete Maria
author_sort Araldi, Rodrigo Pinheiro
collection PubMed
description Although papillary thyroid carcinoma (PTC) has a good prognosis, 20–90% of patients show metastasis to regional lymph nodes and 10–15% of patients show metastasis to distant sites. Metastatic disease represents the main clinical challenge that impacts survival rate. We previously showed that LIMD2 was a novel metastasis-associated gene. In this study, to interrogate the role of LIMD2 in cancer invasion and metastasis, we used CRISPR-mediated knockout (KO) of LIMD2 in PTC cells (BCPAP and TPC1). Western blot and high-content screening (HCS) analysis confirmed functional KO of LIMD2. LIMD2 KO reduced in vitro invasion and migration. Ultrastructural analyses showed that cell polarity and mitochondria function and morphology were restored in LIMD2 KO cells. To unveil the signals supervising these phenotypic changes, we employed phospho-protein array. Several members of the MAPK superfamily showed robust reduction in phosphorylation. A Venn diagram displayed the overlap of kinases with reduced phosphorylation in both cell lines and showed that they were able to initiate or sustain the epithelial-mesenchymal transition (EMT) and DNA damage checkpoint. Flow cytometry and HCS validation analyses further corroborated the phospho-protein array data. Collectively, our findings show that LIMD2 enhances phosphorylation of kinases associated with EMT and invasion. Through cooperation with different kinases, it contributes to the increased genomic instability that ultimately promotes PTC progression.
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spelling pubmed-77005342020-11-30 LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk Araldi, Rodrigo Pinheiro de Melo, Thatiana Correa Levy, Débora de Souza, Dener Madeiro Maurício, Beatriz Colozza-Gama, Gabriel Avelar Bydlowski, Sergio Paulo Peng, Hongzhuang Rauscher, Frank J. Cerutti, Janete Maria Cells Article Although papillary thyroid carcinoma (PTC) has a good prognosis, 20–90% of patients show metastasis to regional lymph nodes and 10–15% of patients show metastasis to distant sites. Metastatic disease represents the main clinical challenge that impacts survival rate. We previously showed that LIMD2 was a novel metastasis-associated gene. In this study, to interrogate the role of LIMD2 in cancer invasion and metastasis, we used CRISPR-mediated knockout (KO) of LIMD2 in PTC cells (BCPAP and TPC1). Western blot and high-content screening (HCS) analysis confirmed functional KO of LIMD2. LIMD2 KO reduced in vitro invasion and migration. Ultrastructural analyses showed that cell polarity and mitochondria function and morphology were restored in LIMD2 KO cells. To unveil the signals supervising these phenotypic changes, we employed phospho-protein array. Several members of the MAPK superfamily showed robust reduction in phosphorylation. A Venn diagram displayed the overlap of kinases with reduced phosphorylation in both cell lines and showed that they were able to initiate or sustain the epithelial-mesenchymal transition (EMT) and DNA damage checkpoint. Flow cytometry and HCS validation analyses further corroborated the phospho-protein array data. Collectively, our findings show that LIMD2 enhances phosphorylation of kinases associated with EMT and invasion. Through cooperation with different kinases, it contributes to the increased genomic instability that ultimately promotes PTC progression. MDPI 2020-11-23 /pmc/articles/PMC7700534/ /pubmed/33238381 http://dx.doi.org/10.3390/cells9112522 Text en © 2020 by the authors. Licensee MDPI, Basel, Switzerland. This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution (CC BY) license (http://creativecommons.org/licenses/by/4.0/).
spellingShingle Article
Araldi, Rodrigo Pinheiro
de Melo, Thatiana Correa
Levy, Débora
de Souza, Dener Madeiro
Maurício, Beatriz
Colozza-Gama, Gabriel Avelar
Bydlowski, Sergio Paulo
Peng, Hongzhuang
Rauscher, Frank J.
Cerutti, Janete Maria
LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk
title LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk
title_full LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk
title_fullStr LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk
title_full_unstemmed LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk
title_short LIMD2 Regulates Key Steps of Metastasis Cascade in Papillary Thyroid Cancer Cells via MAPK Crosstalk
title_sort limd2 regulates key steps of metastasis cascade in papillary thyroid cancer cells via mapk crosstalk
topic Article
url https://www.ncbi.nlm.nih.gov/pmc/articles/PMC7700534/
https://www.ncbi.nlm.nih.gov/pubmed/33238381
http://dx.doi.org/10.3390/cells9112522
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