The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells

Doppel (Dpl) protein is the paralogue of the cellular prion (PrPC) protein. In humans, Dpl is expressed almost exclusively in testis where it is involved in spermatogenesis. Recently, the protein has been described to be ectopically expressed in astrocytomas and its potential association to the brai...

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Main Authors: Alberto Azzalin, Elena Sbalchiero, Giulia Barbieri, Silvia Palumbo, Cristina Muzzini, Sergio Comincini
Format: Article
Language:English
Published: Wiley 2008-01-01
Series:Cellular Oncology
Online Access:http://dx.doi.org/10.3233/CLO-2008-0437
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author Alberto Azzalin
Elena Sbalchiero
Giulia Barbieri
Silvia Palumbo
Cristina Muzzini
Sergio Comincini
author_facet Alberto Azzalin
Elena Sbalchiero
Giulia Barbieri
Silvia Palumbo
Cristina Muzzini
Sergio Comincini
author_sort Alberto Azzalin
collection DOAJ
description Doppel (Dpl) protein is the paralogue of the cellular prion (PrPC) protein. In humans, Dpl is expressed almost exclusively in testis where it is involved in spermatogenesis. Recently, the protein has been described to be ectopically expressed in astrocytomas and its potential association to the brain tumor malignancy progression has been advanced. In this study, we aimed to investigate in vitro the potential involvement of Dpl in the tumor cell migration: to this purpose, Dpl expression was reduced in the IPDDC-A2 astrocytoma-derived cell line, by means of antisense and siRNA approaches; migration rates were then evaluated by means of a scratch wound healing assay. As a result, the cellular migration was sensibly reduced after Dpl silencing. Following a complementary approach, in HeLa cells, showing very low endogenous Dpl expression, the protein expression was induced by transfection and stabilization of an eukaryotic expression vector containing the doppel gene coding sequence. These stably Dpl-overexpressing cells revealed a significant increase in the migration rate, compared to untreated and control cells. In addition, Dpl-forced expression induced substantial changes in the cell morphology. Of note, in these cells, viability examination by means of tetrazolium-based assay did not reveal differences in the proliferation; on the contrary, a variation in density-dependent growth, leading to an increase of cell contact inhibition was highlighted. These results, in conclusion, might suggest a potential and functional role for Dpl in tumor cells migratory and morphological behaviours and address to future gene-targeted therapeutic interventions.
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spelling doaj-art-8dc0b8419a65456fbc986072191211432025-02-03T05:47:49ZengWileyCellular Oncology1570-58701875-86062008-01-0130649150110.3233/CLO-2008-0437The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived CellsAlberto Azzalin0Elena Sbalchiero1Giulia Barbieri2Silvia Palumbo3Cristina Muzzini4Sergio Comincini5Dipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ItalyDipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ItalyDipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ItalyDipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ItalyDipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ItalyDipartimento di Genetica e Microbiologia, Università di Pavia, Pavia, ItalyDoppel (Dpl) protein is the paralogue of the cellular prion (PrPC) protein. In humans, Dpl is expressed almost exclusively in testis where it is involved in spermatogenesis. Recently, the protein has been described to be ectopically expressed in astrocytomas and its potential association to the brain tumor malignancy progression has been advanced. In this study, we aimed to investigate in vitro the potential involvement of Dpl in the tumor cell migration: to this purpose, Dpl expression was reduced in the IPDDC-A2 astrocytoma-derived cell line, by means of antisense and siRNA approaches; migration rates were then evaluated by means of a scratch wound healing assay. As a result, the cellular migration was sensibly reduced after Dpl silencing. Following a complementary approach, in HeLa cells, showing very low endogenous Dpl expression, the protein expression was induced by transfection and stabilization of an eukaryotic expression vector containing the doppel gene coding sequence. These stably Dpl-overexpressing cells revealed a significant increase in the migration rate, compared to untreated and control cells. In addition, Dpl-forced expression induced substantial changes in the cell morphology. Of note, in these cells, viability examination by means of tetrazolium-based assay did not reveal differences in the proliferation; on the contrary, a variation in density-dependent growth, leading to an increase of cell contact inhibition was highlighted. These results, in conclusion, might suggest a potential and functional role for Dpl in tumor cells migratory and morphological behaviours and address to future gene-targeted therapeutic interventions.http://dx.doi.org/10.3233/CLO-2008-0437
spellingShingle Alberto Azzalin
Elena Sbalchiero
Giulia Barbieri
Silvia Palumbo
Cristina Muzzini
Sergio Comincini
The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells
Cellular Oncology
title The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells
title_full The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells
title_fullStr The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells
title_full_unstemmed The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells
title_short The doppel (Dpl) Protein Influences In Vitro Migration Capability in Astrocytoma-Derived Cells
title_sort doppel dpl protein influences in vitro migration capability in astrocytoma derived cells
url http://dx.doi.org/10.3233/CLO-2008-0437
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