Clinical & Experimental Metastasis

, Volume 31, Issue 4, pp 461–474 | Cite as

The reciprocal interactions between astrocytes and prostate cancer cells represent an early event associated with brain metastasis

  • Eliane Gouvêa de Oliveira Barros
  • Antonio PalumboJr.
  • Pedro Lucas Prado Mello
  • Rômulo Medina de Mattos
  • Julianna Henriques da Silva
  • Bruno Pontes
  • Nathan Bessa Viana
  • Rackele Ferreira do Amaral
  • Flavia Regina Souza Lima
  • Nathalia Meireles da Costa
  • Celia Yelimar Palmero
  • Leandro Miranda-Alves
  • Christina Maeda Takiya
  • Luiz Eurico Nasciutti
Article

Abstract

Tumor establishment, growth, and survival are supported by interactions with microenvironment components. Here, we investigated whether the interactions between prostate cancer cells and cortical astrocytes are associated to a potential role for astrocytes in tumor establishment. We demonstrate that astrocytes interact in vitro with prostatic cancers cells derived from different metastatic sites. Astrocytes and their secreted extracellular matrix, stimulate DU145 cell (a brain-derived prostate tumor cell line) proliferation while inhibiting cell death and modulating the expression of several genes related to prostate cancer progression, suggesting the activation of EMT process in these cells. In contrast, DU145 cells and their conditioned medium inhibited cell proliferation and induced cell death of astrocytes. On the other hand, the astrocytes were unable to significantly induce an increment of LNCaP cell (a lymph node-derived prostate tumor cell line) proliferative activity. In addition, LNCaP cells were also unable to induce cell death of astrocytes. Thus, we believe that DU145 cells, but not LNCaP cells, present an even more aggressive behavior when interacting with astrocytes. These results provide an important contribution to the elucidation of the cellular mechanisms involved in the brain microenvironment colonization.

Keywords

Prostate cancer Brain metastasis Astrocytes DU145 cells LNCaP cells Epithelial mesenchymal transition TGFβ 

Abbreviations

EMT

Epithelial mesenchymal transition

TGF-β

Transforming growth factor beta

CNS

Central nervous system

ECM

Extracellular matrix

FBS

Fetal bovine serum

24-CM

Conditioned medium collected after 24 h of culture

48-CM

Conditioned medium collected after 48 h of culture

PBS

Phosphate buffered saline

GFAP

Glial fibrillary acidic protein

CK

Cytokeratin

MTT

3-(4,5-Dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide

VT

Tangential velocity

VS

Spreading velocity

MMP

Matrix metalloproteinase

Supplementary material

10585_2014_9640_MOESM1_ESM.tif (5.7 mb)
Electronic Supplementary Material Fig. 1 Characterization by immunofluorescence and cell proliferation and cell death analysis of astrocytes and LNCaP cells in co-culture. (A)The cells were incubated for 24 and 48 h in medium supplemented with 0.5 FBS, and immunostained with anti-GFAP and human anti-cytokeratin. The astrocytes (red) LNCaP cells (white). In co-culture, LNCaP cells formed small clusters arranged over the astrocytes monolayer. (B) Quantification of cell number present in co-culture by flow cytometry. (C) TUNEL assay performed in co-culture of astrocytes and LNCaP cells. DAPI (blue), nuclei staining. Scale bar, 200 µm (TIFF 5792 kb)
10585_2014_9640_MOESM2_ESM.tif (2.1 mb)
Electronic Supplementary Material Fig. 2 Cell viability analysis of astrocytes and DU145 cells isolated and in co-culture. Cell viability was measured by the MTT colorimetric assay after 24 (A) and 48 (B) h of culture in supplemented medium with 10 %, 0.5 %, and without FBS. Note that after 48 h in 0.5 % FBS, the viability of astrocytes begins to be rescue. The results are expressed as the means ± CI of three independent experiments. CI = confidence interval. (*)p < 0.05, (**) p < 0.01, (***) p < 0.001 (TIFF 2144 kb)
10585_2014_9640_MOESM3_ESM.tif (2.2 mb)
Electronic Supplementary Material Fig. 3 Schematic model of co-culture and cell sorting. (A) Schematic model of co-culture of astrocytes labeled with Green CMFDA and unlabeled DU145 cells. (B) Separation of cells by flow cytometry. Note that there are differences in fluorescence emission peaks of labeled astrocytes and unlabeled DU145 cells (TIFF 2228 kb)

Supplementary material 4 (MPG 1378 kb)

10585_2014_9640_MOESM5_ESM.docx (34 kb)
Supplementary material 5 (DOCX 34 kb)

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Copyright information

© Springer Science+Business Media Dordrecht 2014

Authors and Affiliations

  • Eliane Gouvêa de Oliveira Barros
    • 1
  • Antonio PalumboJr.
    • 1
  • Pedro Lucas Prado Mello
    • 1
  • Rômulo Medina de Mattos
    • 1
  • Julianna Henriques da Silva
    • 1
  • Bruno Pontes
    • 1
    • 2
  • Nathan Bessa Viana
    • 2
    • 3
  • Rackele Ferreira do Amaral
    • 1
  • Flavia Regina Souza Lima
    • 1
  • Nathalia Meireles da Costa
    • 4
  • Celia Yelimar Palmero
    • 1
  • Leandro Miranda-Alves
    • 1
  • Christina Maeda Takiya
    • 5
  • Luiz Eurico Nasciutti
    • 1
  1. 1.Programa de Pesquisa em Biologia Celular e do Desenvolvimento, Instituto de Ciências BiomédicasUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  2. 2.LPO/COPEA, Instituto de Ciências BiomédicasUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  3. 3.Instituto de FísicaUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil
  4. 4.Programa de Carcinogênese Molecular, Centro de Pesquisas (CPQ)Instituto Nacional de Câncer (INCA)Rio de JaneiroBrazil
  5. 5.Instituto de Biofísica Carlos Chagas FilhoUniversidade Federal do Rio de JaneiroRio de JaneiroBrazil

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