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Functional analysis of dual-specificity protein phosphatases in angiogenesis
Amand, Mathieu; ERPICUM, Charlotte; Gilles, Christine et al.
2016In Pulido, Rafael (Ed.) Protein Tyrosine Phosphatases: Methods and Protocols
Peer reviewed
 

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Abstract :
[en] Therapeutic perspectives targeting angiogenesis in cancer stimulated an intense investigation of the mechanisms triggering and governing angiogenic processes. Several publications have highlighted the importance of typical dual-specificity phosphatases (DSPs) or MKPs in endothelial cells and their role in controlling different biological functions implicated in angiogenesis such as migration, proliferation, apoptosis, tubulogenesis and cell adhesion. However, among atypical DSPs, the only one investigated in angiogenesis was DUSP3. We recently identified this DSP as new key player in endothelial cells and angiogenesis. In this chapter we provide with detailed protocols and models used to investigate the role of DUSP3 in endothelial cells and angiogenesis. We start the chapter with an overview of the role of several DSPs in angiogenesis. We continue with providing a full description of a highly efficient transfection protocol to deplete DUSP3 using small interfering RNA (siRNA) in the primary Human Umbilical Vein Endothelial Cells (HUVEC). We next describe the major assays used to investigate different processes involved in angiogenesis such as tube formation assay, proliferation assay and spheroids sprouting assay. We finish the chapter by validating our results in DUSP3-knockout mice using in vivo angiogenesis assays such as Matrigel plug and Lewis lung carcinoma cell subcutaneous xenograft model followed by anti-CD31 immunofluorescence and ex vivo aortic ring assay. All methods described can be adapted to other phosphatases and signaling molecules.
Disciplines :
Oncology
Author, co-author :
Amand, Mathieu
ERPICUM, Charlotte ;  Centre Hospitalier Universitaire de Liège - CHU > Anatomie pathologique
Gilles, Christine ;  Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
Noël, Agnès ;  Université de Liège > Département des sciences cliniques > Labo de biologie des tumeurs et du développement
Rahmouni, Souad  ;  Université de Liège > Département des sciences cliniques > GIGA-R:Immunopath. - Maladies infect. et médec. inter. gén.
Language :
English
Title :
Functional analysis of dual-specificity protein phosphatases in angiogenesis
Publication date :
12 August 2016
Main work title :
Protein Tyrosine Phosphatases: Methods and Protocols
Editor :
Pulido, Rafael
Publisher :
Springer Science, New York, United States
Pages :
331-49
Peer reviewed :
Peer reviewed
Available on ORBi :
since 23 November 2015

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