Downregulation of uPAR promotes urokinase translocation into the nucleus and epithelial to mesenchymal transition in neuroblastoma

Autor: Semina, Ekaterina V., Rubina, Kseniya A., Shmakova, Anna A., Rysenkova, Karina D., Klimovich, Polina S., Aleksanrushkina, Natalya A., Sysoeva, Veronika Y., Karagyaur, Maxim N., Tkachuk, Vsevolod A.
Rok vydání: 2020
Předmět:
0301 basic medicine
Epithelial-Mesenchymal Transition
Physiology
Plasmin
Clinical Biochemistry
urokinase
epithelial–mesenchymal transition
Receptors
Urokinase Plasminogen Activator
Gene Knockout Techniques
Neuroblastoma
03 medical and health sciences
0302 clinical medicine
Downregulation and upregulation
Original Research Articles
Cell Line
Tumor
urokinase receptor
medicine
Humans
Original Research Article
Epithelial–mesenchymal transition
skin and connective tissue diseases
CRISPR/Cas9
neoplasms
Transcription factor
Cell Nucleus
Chemistry
Membrane Proteins
Cell migration
Cell Biology
biological factors
Cell biology
Gene Expression Regulation
Neoplastic
Urokinase receptor
030104 developmental biology
030220 oncology & carcinogenesis
CRISPR-Cas Systems
biological phenomena
cell phenomena
and immunity
Signal transduction
Plasminogen activator
Signal Transduction
medicine.drug
Zdroj: Journal of Cellular Physiology
ISSN: 1097-4652
0021-9541
Popis: The urokinase system is involved in a variety of physiological processes, such as fibrinolysis, matrix remodeling, wound healing, and regeneration. Upon binding to its cognate receptor urokinase‐type plasminogen activator receptor (uPAR), urokinase‐type plasminogen activator (uPA) catalyzes the conversion of plasminogen to plasmin and the activation of matrix metalloproteases. Apart from this, uPA–uPAR interaction can lead to the activation of transcription factors, mitogen‐activated protein kinase signaling pathways and RTK cascades. Elevated expression of uPA and uPAR is markedly associated with cancer progression and metastasis and correlates with a poor prognosis in clinics. Targeting the urokinase system has proved to be effective in experimental models in vitro and in vivo, however, in clinics the inhibition of the uPA/uPAR system has fallen short of expectations, suggesting that the question of the functional relevance of uPA/uPAR system is far from being moot. Recently, using CRISPR/Cas9 technology, we have shown that uPAR knockout decreases the proliferation of neuroblastoma Neuro2a cells in vitro. In the present study we demonstrate that uPAR expression is essential for maintaining the epithelial phenotype in Neuro2a cells and that uPAR silencing promotes epithelial‐mesenchymal transition (EMT) and increased cell migration. Accordingly, uPAR knockout results in the downregulation of epithelial markers (E‐cadherin, occludin, and claudin‐5) and in the increase of mesenchymal markers (N‐cadherin, α‐smooth muscle actin, and interleukin‐6). In search of the molecular mechanism underlying these changes, we identified uPA as a key component. Two key insights emerged as a result of this work: in the absence of uPAR, uPA is translocated into the nucleus where it is presumably involved in the activation of transcription factors (nuclear factor κB and Snail) resulting in EMT. In uPAR‐expressing cells, uPAR functions as a uPA “trap” that binds uPA on the cell surface and promotes controlled uPA internalization and degradation in lysosomes.
We found that extracellular protease urokinase‐type plasminogen activator (uPA), in the absence of its receptor urokinase‐type plasminogen activator receptor (uPAR), is translocated into the nucleus where it is involved in the activation of transcription factors (nuclear factor κB and Snail) resulting in epithelial–mesenchymal transition, decreased adhesion and increased migration of neuroblastoma cells. Consistently, in uPAR‐expressing cells, uPAR functions as a uPA “trap” that binds uPA on the cell surface and promotes controlled uPA internalization and degradation in lysosomes.
Databáze: OpenAIRE
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