HeberFERON, a co-formulation of Interferon (IFN)-α2b and IFN-γ, has effects on skin cancer and other solid tumors. It has antiproliferative effects over glioblastoma multiform (GBM) clones and cultured cell lines, including U-87 MG. Here, we report the first label-free quantitative proteomic and phospho-proteomic analyses to evaluate changes induced by HeberFERON after 72 h incubation of U-87 MG that can explain the effect on cellular proliferation. LC-MS/MS, functional enrichment and networking analysis were performed. We identified 7627 proteins; 122 and 211 were down- and up-regulated by HeberFERON (fold change > 2; p < 0.05), respectively. We identified 23,549 peptides (5692 proteins) and 8900 phospho-peptides; 523 of these phospho-peptides (359 proteins) were differentially modified. Proteomic enrichment showed IFN signaling and its control, direct and indirect antiviral mechanisms were the main modulated processes. Phospho-proteome enrichment displayed the cell cycle as one of the most commonly targeted events together with cytoskeleton organization; translation/RNA splicing, autophagy and DNA repair, as represented biological processes. There is a high interconnection of phosphoproteins in a molecular network; mTOR occupies a centric hub with interactions with translation machinery, cytoskeleton and autophagy components. Novel phosphosites and others with unknown biological functionality in key players in the aforementioned processes were regulated by HeberFERON and involved CDK and ERK kinases. These findings open new experimental hypotheses regarding HeberFERON action. The results obtained contribute to a better understanding of HeberFERON effector mechanisms in the context of GBM treatment.
HeberFERON is a co-formulation of Interferon (IFN)-alpha2b and IFN-gamma; in synergic proportions, with a demonstrated effect on skin cancer and other solid tumors. It has antiproliferative effects over glioblastoma multiform (GBM) clones and cell lines in culture, including U-87 MG. Omics studies in U-87 MG showed distinctive expression patterns compared to individual IFNs. Kinase signaling pathways dysregulation can also contribute to HeberFERON effects. Here, we report the first label-free quantitative proteomic and phosphoproteomic analyses to evaluate changes induced by HeberFERON after 72h incubation of U-87 MG cell line. LC-MS/MS analysis identified 7627 proteins with a fold change >2 (p<0.05); 122 and 211 were down- and up-regulated by HeberFERON, respectively. We identified 23549 peptides (5692 proteins) and 8900 phosphopeptides, 412 of these phosphopeptides (359 proteins) were differentially modified with fold change >2 (p<0.05). Proteomic enrichment analysis showed IFN signaling and its control, together to direct and indirect antiviral mechanisms were the main modulated processes. Enrichment analysis of phosphoproteome pointed to the cell cycle, cytoskeleton organization, translation and RNA splicing, autophagy, and DNA repair as biological processes represented. There is a high interconnection of phosphoproteins in a molecular network, where mTOR occupies a centric hub. HeberFERON regulates many phosphosites newly reported or with no clear association to kinases. Of interest is phosphosites increasing phosphorylation were mainly modified by CDK and ERK kinases, thus new cascades regulations can be determining the antiproliferation outcome. Our results contribute to a better mechanistic understanding of HeberFERON in the context of GBM.
scite is a Brooklyn-based organization that helps researchers better discover and understand research articles through Smart Citations–citations that display the context of the citation and describe whether the article provides supporting or contrasting evidence. scite is used by students and researchers from around the world and is funded in part by the National Science Foundation and the National Institute on Drug Abuse of the National Institutes of Health.
customersupport@researchsolutions.com
10624 S. Eastern Ave., Ste. A-614
Henderson, NV 89052, USA
Copyright © 2024 scite LLC. All rights reserved.
Made with 💙 for researchers
Part of the Research Solutions Family.