TY - JOUR
T1 - Glutaminase isoforms expression switches microRNA levels and oxidative status in glioblastoma cells
AU - de los Santos-Jiménez, Juan
AU - Campos-Sandoval, José A.
AU - Márquez-Torres, Clara
AU - Urbano-Polo, Nieves
AU - Brøndegaard, David
AU - Martín-Rufián, Mercedes
AU - Lobo, Carolina
AU - Peñalver, Ana
AU - Gómez-García, María C.
AU - Martín-Campos, Janet
AU - Cardona, Carolina
AU - Castilla, Laura
AU - da Costa Souza, Felipe
AU - Cheng, Tzuling
AU - Segura, Juan A.
AU - Alonso, Francisco J.
AU - Curi, Rui
AU - Colquhoun, Alison
AU - DeBerardinis, Ralph J.
AU - Márquez, Javier
AU - Matés, José M.
N1 - Funding Information:
This work was financially supported by Ministerio de Ciencia y Tecnología of Spain, RTI2018-096866-B-I00 (to JMM and JM) and Ministerio de Educación of Spain PHB2010-0014-PC (to JMM). RJD is supported by the Howard Hughes Medical Institute, the National Cancer Institute, the Cancer Prevention and Research Institute of Texas, and the Moody Foundation. Thanks are also due to CAPES/DGU 250/11, Brazil (to FCS, RC and AC). JDSJ is granted by FPU17/04084, Ministerio de Ciencia, Innovación y Universidades.
Funding Information:
Thanks are due to Monika Szeliga, Department of Neurotoxicology, Mossakowski Medical Research Centre, Polish Academy of Sciences, Warsaw, Poland. She kindly provided LN229-GAB(+) GBM cell model.
Publisher Copyright:
© 2021, The Author(s).
PY - 2021/12
Y1 - 2021/12
N2 - Background: Glutaminase isoenzymes GLS and GLS2 play apparently opposing roles in cancer: GLS acts as an oncoprotein, while GLS2 (GAB isoform) has context specific tumour suppressive activity. Some microRNAs (miRNAs) have been implicated in progression of tumours, including gliomas. The aim was to investigate the effect of GLS and GAB expression on both miRNAs and oxidative status in glioblastoma cells. Methods: Microarray profiling of miRNA was performed in GLS-silenced LN229 and GAB-transfected T98G human glioblastoma cells and their wild-type counterparts. Results were validated by real-time quantitative RT-PCR. Oxidative status and antioxidant enzymes were determined by spectrophotometric or fluorescence assays in GLS-silenced LN229 and T98G, and GAB-transfected LN229 and T98G. Results: MiRNA-146a-5p, miRNA-140-3p, miRNA-21-5p, miRNA-1260a, and miRNA-92a-3p were downregulated, and miRNA-1246 was upregulated when GLS was knocked down. MiRNA-140-3p, miRNA-1246, miRNA-1260a, miRNA-21-5p, and miRNA-146a-5p were upregulated when GAB was overexpressed. Oxidative status (lipid peroxidation, protein carbonylation, total antioxidant capacity, and glutathione levels), as well as antioxidant enzymes (catalase, superoxide dismutase, and glutathione reductase) of silenced GLS glioblastoma cells and overexpressed GAB glioblastoma cells significantly changed versus their respective control glioblastoma cells. MiRNA-1246, miRNA-1260a, miRNA-146a-5p, and miRNA-21-5p have been characterized as strong biomarkers of glioblastoma proliferation linked to both GLS silencing and GAB overexpression. Total glutathione is a reliable biomarker of glioblastoma oxidative status steadily associated to both GLS silencing and GAB overexpression. Conclusions: Glutaminase isoenzymes are related to the expression of some miRNAs and may contribute to either tumour progression or suppression through certain miRNA-mediated pathways, proving to be a key tool to switch cancer proliferation and redox status leading to a less malignant phenotype. Accordingly, GLS and GAB expression are especially involved in glutathione-dependent antioxidant defence.
AB - Background: Glutaminase isoenzymes GLS and GLS2 play apparently opposing roles in cancer: GLS acts as an oncoprotein, while GLS2 (GAB isoform) has context specific tumour suppressive activity. Some microRNAs (miRNAs) have been implicated in progression of tumours, including gliomas. The aim was to investigate the effect of GLS and GAB expression on both miRNAs and oxidative status in glioblastoma cells. Methods: Microarray profiling of miRNA was performed in GLS-silenced LN229 and GAB-transfected T98G human glioblastoma cells and their wild-type counterparts. Results were validated by real-time quantitative RT-PCR. Oxidative status and antioxidant enzymes were determined by spectrophotometric or fluorescence assays in GLS-silenced LN229 and T98G, and GAB-transfected LN229 and T98G. Results: MiRNA-146a-5p, miRNA-140-3p, miRNA-21-5p, miRNA-1260a, and miRNA-92a-3p were downregulated, and miRNA-1246 was upregulated when GLS was knocked down. MiRNA-140-3p, miRNA-1246, miRNA-1260a, miRNA-21-5p, and miRNA-146a-5p were upregulated when GAB was overexpressed. Oxidative status (lipid peroxidation, protein carbonylation, total antioxidant capacity, and glutathione levels), as well as antioxidant enzymes (catalase, superoxide dismutase, and glutathione reductase) of silenced GLS glioblastoma cells and overexpressed GAB glioblastoma cells significantly changed versus their respective control glioblastoma cells. MiRNA-1246, miRNA-1260a, miRNA-146a-5p, and miRNA-21-5p have been characterized as strong biomarkers of glioblastoma proliferation linked to both GLS silencing and GAB overexpression. Total glutathione is a reliable biomarker of glioblastoma oxidative status steadily associated to both GLS silencing and GAB overexpression. Conclusions: Glutaminase isoenzymes are related to the expression of some miRNAs and may contribute to either tumour progression or suppression through certain miRNA-mediated pathways, proving to be a key tool to switch cancer proliferation and redox status leading to a less malignant phenotype. Accordingly, GLS and GAB expression are especially involved in glutathione-dependent antioxidant defence.
KW - Antioxidant enzymes
KW - Cancer
KW - Glioblastoma
KW - Glutaminase
KW - Oxidative stress
KW - microRNA
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U2 - 10.1186/s12929-021-00712-y
DO - 10.1186/s12929-021-00712-y
M3 - Article
C2 - 33610185
AN - SCOPUS:85101120972
SN - 1021-7770
VL - 28
JO - Journal of Biomedical Science
JF - Journal of Biomedical Science
IS - 1
M1 - 14
ER -