Study of negative feedback between wild-type BRAF or RAFV600E and Mps1 in melanoma / 中华病理学杂志
Chinese Journal of Pathology
;
(12): 274-277, 2015.
Artigo
em Chinês
| WPRIM
| ID: wpr-298119
ABSTRACT
<p><b>OBJECTIVE</b>To study the effect of Mps1 on BRAFWT/MEK/ERK pathway in the presence of wild type BRAF or BRAFV600E in melanoma.</p><p><b>METHODS</b>Melanoma cells harboring BRAFWT genotype were transfected either with pBabe-puro-GST-BRAF-WT and/or pBabe-puro-GFP-Mps1-WT or pBabe-puro-GST-BRAFV600E and/or pBabe-puro-GFP-Mps1-WT, followed by Western blot to detect Mps1 and p-ERK expression. The melanoma cells harboring BRAFWT and BRAFV600E genotype were infected with pSUPER-Mps1 retrovirus to knockdown the endogenous Mps1 protein, followed by Western blot to detect Mps1 and p-ERK expression. Meanwhile, melanoma cells harboring BRAFV600E genotype were infected with pBabe-puro-GFP-Mps1 and Western blot was performed to detect Mps1 and p-ERK expression.</p><p><b>RESULTS</b>In melanoma cells harboring BRAFWT genotype and transfected with pBabe-puro-GST-BRAF-WT and pBabe-puro-GFP-Mps1-WT, phospho-ERK levels were notably reduced as compared to either negative control or empty vector. However, cells transfected with pBabe-puro-GST-BRAFV600E and pBabe-puro-GFP-Mps1-WT, phospho-ERK levels did not change significantly compared with either negative control or empty vector. Knockout of Mps1 in BRAF wild-type cell lines led to an increased ERK activity. However, there was no significant change of ERK activity in BRAFV600E cell lines in the absence of Mps1. The expression of p-ERK in BRAFV600E mutant cell lines infected with pBabe-puro-GFP-Mps1-WT did not show any significant difference from either negative control or empty vector.</p><p><b>CONCLUSIONS</b>Based on these findings, it suggests that there exists an auto-regulatory negative feedback loop between the Mps1 kinase and BRAFWT/ERK signaling. Oncogenic BRAFV600E abrogates the regulatory negative feedback loop of Mps1 on the MAPK pathway.</p>
Texto completo:
DisponíveL
Índice:
WPRIM (Pacífico Ocidental)
Assunto principal:
Fenótipo
/
Proteínas Tirosina Quinases
/
Transfecção
/
Transdução de Sinais
/
Proteínas Serina-Treonina Quinases
/
Proteínas de Ciclo Celular
/
Sistema de Sinalização das MAP Quinases
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Linhagem Celular Tumoral
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Proteínas Proto-Oncogênicas B-raf
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Genética
Limite:
Humanos
Idioma:
Chinês
Revista:
Chinese Journal of Pathology
Ano de publicação:
2015
Tipo de documento:
Artigo
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