MEK Inhibition Reverses Aberrant Signaling in Melanoma Cells through Reorganization of NRas and BRAF in Self Nanoclusters.
Cell Line, Tumor
Cell Membrane
/ drug effects
Epidermal Growth Factor
/ pharmacology
Extracellular Signal-Regulated MAP Kinases
/ metabolism
GTP Phosphohydrolases
/ genetics
Humans
MAP Kinase Kinase 1
/ antagonists & inhibitors
Melanoma
/ drug therapy
Membrane Proteins
/ genetics
Mutation
Protein Kinase Inhibitors
/ pharmacology
Proto-Oncogene Proteins B-raf
/ genetics
Pyridones
/ pharmacology
Pyrimidinones
/ pharmacology
Signal Transduction
/ drug effects
Single Molecule Imaging
Skin Neoplasms
/ drug therapy
Melanoma, Cutaneous Malignant
Journal
Cancer research
ISSN: 1538-7445
Titre abrégé: Cancer Res
Pays: United States
ID NLM: 2984705R
Informations de publication
Date de publication:
01 03 2021
01 03 2021
Historique:
received:
12
04
2020
revised:
29
07
2020
accepted:
14
12
2020
pubmed:
24
12
2020
medline:
21
5
2021
entrez:
23
12
2020
Statut:
ppublish
Résumé
Hotspot mutations of the oncogenes BRAF and NRas are the most common genetic alterations in cutaneous melanoma. Still, the nanoscale organization and signal coupling of these proteins remain incompletely understood, particularly upon expression of oncogenic NRas mutants. Here we employed single-molecule localization microscopy to study the nanoscale organization of NRas and BRAF at the plasma membrane (PM) of melanoma cells. NRas and BRAF resided in self-clusters that did not associate well in resting cells. In EGF-activated cells, NRas clusters became more diffused while overall protein levels at the PM increased; thus allowing enhanced association of NRas and BRAF and downstream signaling. In multiple melanoma cell lines, mutant NRas resided in more pronounced self-clusters relative to wild-type (WT) NRas yet associated more with the clustered and more abundant BRAF. In cells resistant to trametinib, a clinical MEK inhibitor (MEKi), a similar coclustering of NRas and BRAF was observed upon EGF activation. Strikingly, treatment of cells expressing mutant NRas with trametinib reversed the effect of mutant NRas expression by restoring the nonoverlapping self-clusters of NRas and BRAF and by reducing their PM levels and elevated pERK levels caused by mutant NRas. Our results indicate a new mechanism for signal regulation of NRas in melanoma through its nanoscale dynamic organization and a new mechanism for MEKi function in melanoma cells carrying NRas mutations but lacking MEK mutations. SIGNIFICANCE: Nanoscale dynamic organization of WT and mutant NRas relative to BRAF serves as a regulatory mechanism for NRas signaling and may be a viable therapeutic target for its sensitivity to MEKi.
Identifiants
pubmed: 33355187
pii: 0008-5472.CAN-20-1205
doi: 10.1158/0008-5472.CAN-20-1205
doi:
Substances chimiques
Membrane Proteins
0
Protein Kinase Inhibitors
0
Pyridones
0
Pyrimidinones
0
trametinib
33E86K87QN
Epidermal Growth Factor
62229-50-9
BRAF protein, human
EC 2.7.11.1
Proto-Oncogene Proteins B-raf
EC 2.7.11.1
Extracellular Signal-Regulated MAP Kinases
EC 2.7.11.24
MAP Kinase Kinase 1
EC 2.7.12.2
GTP Phosphohydrolases
EC 3.6.1.-
NRAS protein, human
EC 3.6.1.-
Types de publication
Journal Article
Research Support, Non-U.S. Gov't
Langues
eng
Sous-ensembles de citation
IM
Pagination
1279-1292Informations de copyright
©2020 American Association for Cancer Research.
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