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The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis.

Augustin I, Goidts V, Bongers A, Kerr G, Vollert G, Radlwimmer B, Hartmann C, Herold-Mende C, Reifenberger G, von Deimling A, Boutros M - EMBO Mol Med (2011)

Bottom Line: Malignant astrocytomas are highly aggressive brain tumours with poor prognosis.Evi/Wls is a core Wnt signalling component and a specific regulator of pan-Wnt protein secretion, affecting both canonical and non-canonical signalling.We further show that Evi/Wls overexpression is sufficient to promote downstream Wnt signalling.

View Article: PubMed Central - PubMed

Affiliation: German Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics and Heidelberg University, Faculty of Medicine Mannheim, Department of Cell and Molecular Biology, Heidelberg, Germany. i.augustin@dkfz.de

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Evi overexpressing ESCs showed increased Wnt responseImmunofluorescence of endogeneous expression of Evi-YFP in ESC colonies.Relative mRNA expression levels of Evi were analysed by quantitative RT-PCR. Evi-ESCs expressed increased Evi levels.Western blot of Evi overexpressing ESCs. Evi-ESCs expressed Evi-YFP fusion protein.Evi-ESCs stably transfected with 7TCF Firefly luciferase reporter showed increased Wnt reporter activity (*p < 0.05).Transfection of 7TCF Firefly luciferase reporter containing HEK293 cells with Evi-GFP plasmid led to increased Wnt reporter activity compared to transfection with GFP control vector (**p < 0.01).
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fig02: Evi overexpressing ESCs showed increased Wnt responseImmunofluorescence of endogeneous expression of Evi-YFP in ESC colonies.Relative mRNA expression levels of Evi were analysed by quantitative RT-PCR. Evi-ESCs expressed increased Evi levels.Western blot of Evi overexpressing ESCs. Evi-ESCs expressed Evi-YFP fusion protein.Evi-ESCs stably transfected with 7TCF Firefly luciferase reporter showed increased Wnt reporter activity (*p < 0.05).Transfection of 7TCF Firefly luciferase reporter containing HEK293 cells with Evi-GFP plasmid led to increased Wnt reporter activity compared to transfection with GFP control vector (**p < 0.01).

Mentions: Next, we asked whether Evi overexpression is sufficient to lead to an increase in Wnt signalling. To this end, we generated Evi overexpressing embryonic stem cells (ESCs) by targeted insertion of a C-terminal tagged Evi-YFP into the ROSA26-locus (Fig S3 of Supporting information). Immunofluorescence analysis of ROSA26::Evi-YFP ESC confirmed Evi-YFP fusion protein expression with perinuclear enrichment (Fig 2A). Quantitative RT-PCR revealed elevated Evi transcription in ROSA26::Evi-YFP ESCs and Western blot analysis with an antibody against Evi detected endogenous and Evi-YFP fusion protein (Fig 2B and C).


The Wnt secretion protein Evi/Gpr177 promotes glioma tumourigenesis.

Augustin I, Goidts V, Bongers A, Kerr G, Vollert G, Radlwimmer B, Hartmann C, Herold-Mende C, Reifenberger G, von Deimling A, Boutros M - EMBO Mol Med (2011)

Evi overexpressing ESCs showed increased Wnt responseImmunofluorescence of endogeneous expression of Evi-YFP in ESC colonies.Relative mRNA expression levels of Evi were analysed by quantitative RT-PCR. Evi-ESCs expressed increased Evi levels.Western blot of Evi overexpressing ESCs. Evi-ESCs expressed Evi-YFP fusion protein.Evi-ESCs stably transfected with 7TCF Firefly luciferase reporter showed increased Wnt reporter activity (*p < 0.05).Transfection of 7TCF Firefly luciferase reporter containing HEK293 cells with Evi-GFP plasmid led to increased Wnt reporter activity compared to transfection with GFP control vector (**p < 0.01).
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Related In: Results  -  Collection

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getmorefigures.php?uid=PMC3306557&req=5

fig02: Evi overexpressing ESCs showed increased Wnt responseImmunofluorescence of endogeneous expression of Evi-YFP in ESC colonies.Relative mRNA expression levels of Evi were analysed by quantitative RT-PCR. Evi-ESCs expressed increased Evi levels.Western blot of Evi overexpressing ESCs. Evi-ESCs expressed Evi-YFP fusion protein.Evi-ESCs stably transfected with 7TCF Firefly luciferase reporter showed increased Wnt reporter activity (*p < 0.05).Transfection of 7TCF Firefly luciferase reporter containing HEK293 cells with Evi-GFP plasmid led to increased Wnt reporter activity compared to transfection with GFP control vector (**p < 0.01).
Mentions: Next, we asked whether Evi overexpression is sufficient to lead to an increase in Wnt signalling. To this end, we generated Evi overexpressing embryonic stem cells (ESCs) by targeted insertion of a C-terminal tagged Evi-YFP into the ROSA26-locus (Fig S3 of Supporting information). Immunofluorescence analysis of ROSA26::Evi-YFP ESC confirmed Evi-YFP fusion protein expression with perinuclear enrichment (Fig 2A). Quantitative RT-PCR revealed elevated Evi transcription in ROSA26::Evi-YFP ESCs and Western blot analysis with an antibody against Evi detected endogenous and Evi-YFP fusion protein (Fig 2B and C).

Bottom Line: Malignant astrocytomas are highly aggressive brain tumours with poor prognosis.Evi/Wls is a core Wnt signalling component and a specific regulator of pan-Wnt protein secretion, affecting both canonical and non-canonical signalling.We further show that Evi/Wls overexpression is sufficient to promote downstream Wnt signalling.

View Article: PubMed Central - PubMed

Affiliation: German Cancer Research Center (DKFZ), Division of Signaling and Functional Genomics and Heidelberg University, Faculty of Medicine Mannheim, Department of Cell and Molecular Biology, Heidelberg, Germany. i.augustin@dkfz.de

Show MeSH
Related in: MedlinePlus