Glioma-astrocyte interaction modifies the astrocyte phenotype in a co-culture experimental model

Nicoletta Gagliano, Francesco Costa, Chiara Cossetti, Letizia Pettinari, Rosaria Bassi, Maurizio Chiriva-Internati, Everardo Cobos, Magda Gioia, Stefano Pluchino

Research output: Contribution to journalArticle

Abstract

As the majority of gliomas arise through malignant transformation of astrocytes, we aimed at investigating the interaction between malignant glioma cells and astrocytes in a co-culture experimental model. For this purpose we analyzed the expression of genes and proteins involved in tumor promotion and invasion, such as glial fibrillary acidic protein (GFAP), matrix metalloproteinase-2 (MMP-2), tissue inhibitor of MMP-2 (TIMP-2), transforming growth factor-β1 (TGF-β1), secreted protein acidic and rich in cysteine (SPARC), and connexin 43 (CX43). Co-cultures of human neural stem cell-derived astrocytes and U87 MG astrocytoma cells were performed in a transwell system. Gene expression was evaluated by real-time RT-PCR, and protein analysis was performed by Western blotting, SDS-zymography, and immunofluorescence. GFAP tended to be up-regulated in astrocytes co-cultivated with U87, suggesting a reactive response induced by glioma cells. CX43 mRNA tended to be down- regulated in co-cultured astrocytes, as well as the non-phosphorylated isoform at the protein level. MMP-2 mRNA tended to be up-regulated, and MMP-2 protein levels were significantly increased in astrocytes co-cultivated with U87. TIMP-2 and SPARC mRNA decreased in astrocytes co-cultivated with U87, showing lower expression in glioma cells. By contrast, SPARC protein expression was strongly induced in supernatants of co-cultured astrocytes. TGF-β1 was not modified. Our results suggest that U87 cells elicit phenotype modifications in the neighbouring resident astrocytes very likely mediated by soluble factors. Glioma/ astrocyte interaction could possibly trigger an astrocyte phenotype modification consistent with a malignant transformation, and favouring a more permissive environment for glioma cells invasion.

Original languageEnglish
Pages (from-to)1349-1356
Number of pages8
JournalOncology Reports
Volume22
Issue number6
DOIs
Publication statusPublished - 2009

Fingerprint

Coculture Techniques
Glioma
Astrocytes
Theoretical Models
Phenotype
Matrix Metalloproteinase 2
Proteins
Cysteine
Connexin 43
Glial Fibrillary Acidic Protein
Transforming Growth Factors
Messenger RNA
Tissue Inhibitor of Metalloproteinase-2
Matrix Metalloproteinase Inhibitors
Neural Stem Cells
Astrocytoma
Fluorescent Antibody Technique
Real-Time Polymerase Chain Reaction
Protein Isoforms
Western Blotting

Keywords

  • Astrocytes
  • Connexin 43
  • Glial fibrillary acidic protein
  • Glioblastoma
  • Matrix metalloproteinase-2
  • Secreted protein acidic and rich in cysteine

ASJC Scopus subject areas

  • Cancer Research
  • Oncology

Cite this

Gagliano, N., Costa, F., Cossetti, C., Pettinari, L., Bassi, R., Chiriva-Internati, M., ... Pluchino, S. (2009). Glioma-astrocyte interaction modifies the astrocyte phenotype in a co-culture experimental model. Oncology Reports, 22(6), 1349-1356. https://doi.org/10.3892/or_00000574

Glioma-astrocyte interaction modifies the astrocyte phenotype in a co-culture experimental model. / Gagliano, Nicoletta; Costa, Francesco; Cossetti, Chiara; Pettinari, Letizia; Bassi, Rosaria; Chiriva-Internati, Maurizio; Cobos, Everardo; Gioia, Magda; Pluchino, Stefano.

In: Oncology Reports, Vol. 22, No. 6, 2009, p. 1349-1356.

Research output: Contribution to journalArticle

Gagliano, N, Costa, F, Cossetti, C, Pettinari, L, Bassi, R, Chiriva-Internati, M, Cobos, E, Gioia, M & Pluchino, S 2009, 'Glioma-astrocyte interaction modifies the astrocyte phenotype in a co-culture experimental model', Oncology Reports, vol. 22, no. 6, pp. 1349-1356. https://doi.org/10.3892/or_00000574
Gagliano, Nicoletta ; Costa, Francesco ; Cossetti, Chiara ; Pettinari, Letizia ; Bassi, Rosaria ; Chiriva-Internati, Maurizio ; Cobos, Everardo ; Gioia, Magda ; Pluchino, Stefano. / Glioma-astrocyte interaction modifies the astrocyte phenotype in a co-culture experimental model. In: Oncology Reports. 2009 ; Vol. 22, No. 6. pp. 1349-1356.
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