Integrative analysis of cell adhesion molecules in glioblastoma identified prostaglandin F2 receptor inhibitor (PTGFRN) as an essential gene.

BACKGROUND: Glioblastoma (GBM) is the most common primary malignant brain tumor in adults exhibiting infiltration into surrounding tissues, recurrence, and resistance to therapy. GBM infiltration is accomplished by many deregulated factors such as cell adhesion molecules (CAMs), which are membrane proteins that participate in cell-cell and cell-ECM interactions to regulate survival, proliferation, migration, and stemness. METHODS: A comprehensive bioinformatics analysis of CAMs (n = 518) in multiple available datasets revealed genetic and epigenetic alterations among CAMs in GBM. Univariate Cox regression analysis using TCGA dataset identified 127 CAMs to be significantly correlated with survival. The poor prognostic indicator PTGFRN was chosen to study its role in glioma. Silencing of PTGFRN in glioma cell lines was achieved by the stable expression of short hairpin RNA (shRNA) against the PTGFRN gene. PTGFRN was silenced and performed cell growth, migration, invasion, cell cycle, and apoptosis assays. Neurosphere and limiting dilution assays were also performed after silencing of PTGFRN in GSCs. RESULTS: Among the differentially regulated CAMs (n = 181, 34.9%), major proportion of them were found to be regulated by miRNAs (n = 95, 49.7%) followed by DNA methylation (n = 32, 16.7%), and gene copy number variation (n = 12, 6.2%). We found that PTGFRN to be upregulated in GBM tumor samples and cell lines with a significant poor prognostic correlation with patient survival. Silencing PTGFRN diminished cell growth, colony formation, anchorage-independent growth, migration, and invasion and led to cell cycle arrest and induction of apoptosis. At the mechanistic level, silencing of PTGFRN reduced pro-proliferative and promigratory signaling pathways such as ERK, AKT, and mTOR. PTGFRN upregulation was found to be due to the loss of its promoter methylation and downregulation of miR-137 in GBM. PTGFRN was also found to be higher in glioma stem-like cells (GSCs) than the matched differentiated glioma cells (DGCs) and is required for GSC growth and survival. Silencing of PTGFRN in GSCs reduced transcript levels of reprogramming factors (Olig2, Pou3f2, Sall2, and Sox2). CONCLUSION: In this study, we provide a comprehensive overview of the differential regulation of CAMs and the probable causes for their deregulation in GBM. We also establish an oncogenic role of PTGFRN and its regulation by miR-137 in GBM, thus signifying it as a potential therapeutic target.

Serine/threonine/tyrosine-interacting-like protein 1 (STYXL1), a pseudo phosphatase, promotes oncogenesis in glioma.

Phosphatases play an important role in cellular signaling and are often found dysregulated in cancers including glioblastoma (GBM). A comprehensive bioinformatics analysis of phosphatases (n = 403) in multiple datasets revealed their deregulation in GBM. Among the differentially regulated phosphatases (n = 186; 46.1%), majority of them were found to be regulated by microRNA (n = 94; 50.5%) followed by DNA methylation (n = 22; 11.8%) and altered copy number variation (n = 10; 5.37%). STYXL1 (Serine/threonine/tyrosine-interacting-like protein 1) was found to be the second most amplified gene in GBM, upregulated, and correlated to poor prognosis. The expression of STYXL1 was also found to be higher in IDH1 mutant gliomas and G-CIMP- gliomas which are reported to be more aggressive than their corresponding counterparts. Silencing STYXL1 inhibited glioma cell growth, soft agar colony formation, migration, invasion, proliferation, and xenograft tumor growth. Further, ectopic expression of STYXL1 was found to promote glioma cell growth, soft agar colony formation, migration, and RasV12 induced in-vitro transformation of immortalized human astrocytes, thus confirming its oncogenic potential in GBM. In this report, we provide a comprehensive overview of deregulation of phosphatases in GBM and demonstrate for the first time, the oncogenic nature of STYXL1 in GBM. This study might be useful for treatment of GBM patients with deregulated STYXL1.