Extracellular adenosine promotes cell migration/invasion of Glioblastoma Stem-like Cells through A₃ Adenosine Receptor activation under hypoxia.

Cancer Lett. 2019 Apr 1;446:112-122. doi: 10.1016/j.canlet.2019.01.004. Epub 2019 Jan 18.

Torres Á¹, Erices JI¹, Sanchez F², Ehrenfeld P³, Turchi L⁴, Virolle T⁴, Uribe D¹, Niechi I¹, Spichiger C¹, Rocha JD¹, Ramirez M⁵, Salazar-Onfray F⁶, San Martín R¹, Quezada C⁷.

Author information

1 Laboratorio de Patología Molecular, Instituto de Bioquímica y Microbiología, Facultad de Ciencias, Universidad Austral de Chile, Valdivia, Chile.
2 Instituto de Inmunología, Universidad Austral de Chile, Valdivia, Chile.
3 Laboratorio de Patología Celular, Instituto de Anatomia, Histología y Patología, Universidad Austral de Chile, Valdivia, Chile; Université Côte d’Azur, Nice, F-06108, France.
4 Université Côte d’Azur, Nice, F-06108, France; CNRS, UMR7277, F-06108, France; Inserm, U1091, Nice, F-06108, France.
5 Servicio de Neurocirugía, Instituto de Neurocirugía Dr. Asenjo, Santiago, Chile; Hospital Clínico Universidad de Chile, Santiago, Chile; Instituto Oncológico Fundación Arturo Lopez Perez (FALP), Santiago, Chile.
6 Instituto Milenio de Inmunología e Inmunoterapia, Facultad de Medicina, Universidad de Chile, Santiago, Chile.
7 Laboratorio de Patología Molecular, Instituto de Bioquímica y Microbiología, Facultad de Ciencias, Universidad Austral de Chile, Valdivia, Chile. Electronic address: claudiaquezada@uach.cl.

Abstract

Glioblastoma (GBM) is the brain tumor with the worst prognosis composed of a cell subpopulation called Glioblastoma Stem-like Cells (GSCs) responsible for tumor recurrence mediated by cell invasion. GSCs persist in a hypoxic microenvironment which promotes extracellular adenosine production and activation of the A₃ Adenosine Receptor (A₃AR), therefore, the aim of this study was to determine the role of extracellular adenosine and A₃AR on GSCs invasion under hypoxia. GSCs were obtained from a U87MG cell line and primary cultures of GBM patients, and then incubated under normoxia or hypoxia. Gene expression was evaluated by RNAseq, RT-qPCR, and western blot. Cell migration was measured by spreading and transwell boyden chamber assays; cell invasion was evaluated by Matrigel-coated transwell, ex vivo brain slice, and in vivo xenograft assays. The contribution of A₃AR on cell migration/invasion was evaluated using the A₃AR antagonist, MRS1220. Extracellular adenosine production was higher under hypoxia than normoxia, mainly by the catalytic action of the prostatic acid phosphatase (PAP), promoting cell migration/invasion in a HIF-2-dependent process. A₃AR blockade decreased cell migration/invasion and the expression of Epithelial-Mesenchymal Transition markers. In conclusion, high levels of extracellular adenosine production enhance cell migration/invasion of GSCs, through HIF-2/PAP-dependent activation of A₃AR under hypoxia.

PMID: 30660649
DOI: 10.1016/j.canlet.2019.01.004