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Influence of NSAIDs and methotrexate on CD73 expression and glioma cell growth

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Abstract

Glioblastoma (GBM) is the most malignant and deadly brain tumor. GBM cells overexpress the CD73 enzyme, which controls the level of extracellular adenosine, an immunosuppressive molecule. Studies have shown that some nonsteroidal anti-inflammatory drugs (NSAIDs) and methotrexate (MTX) have antiproliferative and modulatory effects on CD73 in vitro and in vivo. However, it remains unclear whether the antiproliferative effects of MTX and NSAIDS in GBM cells are mediated by increases in CD73 expression and adenosine formation. The aim of this study was to evaluate the effect of the NSAIDs, naproxen, piroxicam, meloxicam, ibuprofen, sodium diclofenac, acetylsalicylic acid, nimesulide, and ketoprofen on CD73 expression in GBM and mononuclear cells. In addition, we sought to understand whether the effects of MTX may be mediated by CD73 expression and activity. Cell viability and CD73 expression were evaluated in C6 and mononuclear cells after exposure to NSAIDs. For analysis of the mechanism of action of MTX, GBM cells were treated with APCP (CD73 inhibitor), dipyridamole (inhibitor of adenosine uptake), ABT-702 (adenosine kinase enzyme inhibitor), or caffeine (P1 adenosine receptor antagonist), before treatment with MTX and AMP, in the presence or not of mononuclear cells. In summary, only MTX increased the expression of CD73 in GBM cells decreasing cells viability by mechanisms independent of the adenosinergic system. Further studies are needed to understand the role of MTX in the GBM microenvironment.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This study was supported by the following Brazilian agencies: F.F. and A.M.O.B. received support from the Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq/PQ no. 302879/2017-0), Fundação de Amparo à Pesquisa do Estado do Rio Grande do Sul (FAPERGS/PQG numbers: 17/2551- 0000 970-3 and 19/2551-0001783-9 and FAPERGS/PRONEX - project number 16/2551- 0000473-0), and Instituto Nacional de Ciência e Tecnologia (INCT; INCT/CNPq/CAPES/FAPERGS no. 465671/2014-4). A.F.D., L.F.L.S., and J.N.S. were recipients of CNPq fellowship. J.S. received support from the Natural Sciences and Engineering Research Council of Canada (NSERC; RGPIN-2016-05867) and was the recipient of a “Chercheur National” Scholarship from the Fonds de Recherche du Québec – Santé (FRQS).

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Author notes

  1. Daniela Vasconcelos Lopes and Amanda de Fraga Dias contributed equally to this work.

Authors and Affiliations

  1. Departamento de Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Daniela Vasconcelos Lopes, Ana Maria Oliveira Battastini & Fabrício Figueiró

  2. Programa de Pós-Graduação em Ciências Biológicas: Bioquímica, Instituto de Ciências Básicas da Saúde, Universidade Federal do Rio Grande do Sul, Porto Alegre, RS, Brazil

    Amanda de Fraga Dias, Luiz Fernando Lopes Silva, Juliete Nathali Scholl, Ana Maria Oliveira Battastini & Fabrício Figueiró

  3. Département de Microbiologie-Infectiologie et d’Immunologie, Faculté de Médecine, Université Laval, Quebec City, QC, Canada

    Jean Sévigny

  4. Centre de recherche du CHU de Québec-Université Laval, Québec City, QC, G1V 4G2, Canada

    Jean Sévigny

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  1. Daniela Vasconcelos Lopes
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  2. Amanda de Fraga Dias
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  4. Juliete Nathali Scholl
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Contributions

The investigation was planned by D.V.L. and F.F. Cell culture experiments were performed by D.V.L., A.F.D., J.N.S., and L.F.L.S. with guidance from F.F and A.M.O.B. The anti-rat CD73 antibody was donated by J.S. Data processing, and the writing of this manuscript was performed by A.F.D. and L.F.L.S. with input from all co-authors. All authors read and approved the final manuscript.

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Correspondence to Fabrício Figueiró.

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Conflict of interest

Daniela Vasconcelos Lopes declares no competing interests.

Amanda de Fraga Dias declares no competing interests.

Luiz Fernando Lopes Silva declares no competing interests.

Juliete Nathali Scholl declares no competing interests.

Jean Sévigny declares no competing interests.

Ana Maria Oliveira Battastini declares no competing interests.

Fabrício Figueiró declares no competing interests.

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Lopes, D.V., de Fraga Dias, A., Silva, L.F.L. et al. Influence of NSAIDs and methotrexate on CD73 expression and glioma cell growth. Purinergic Signalling 17, 273–284 (2021). https://doi.org/10.1007/s11302-021-09775-w

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