Secretome analysis of Glioblastoma cell line - HNGC-2

Abstract

Glioblastoma multiforme (GBM) is the most common and aggressive type of primary malignant tumor of the central nervous system. We have carried out a deep analysis of the secretome of a rapidly proliferating and tumorigenic cell line HNGC-2, representing GBM, in an effort to identify proteins, which may be targeted in the plasma of GBM patients as markers for diagnosis and disease surveillance. Prefractionation of the proteins from the conditioned medium of HNGC-2 cells in SDS gels followed by LC-MS/MS analysis using an ESI-IT mass spectrometer (LTQ) led to a total of 996 protein identifications with ≥2 peptides each. Of them, 664 proteins were observed in the transcriptome of HNGC-2 cells. The dataset of 996 proteins was mapped to important functional groups, such as cellular assembly and organisation, DNA recombination and repair, and other classes. Actin cytoskeleton signalling, phosphatidyl inositol 3 kinase (PI3K/AKT) and integrin linked kinase (ILK) signalling pathways were seen as enriched pathways. Comparisons with the published secretome of cell lines from 12 different cancers, including GBM, revealed that 348 proteins shared a commonality with a secretome of at least one other cell line, 321 of which were found to contain signal sequences or transmembrane domains and 335 could be linked to a plasma membrane or extracellular localization. Through intergration of this data we arrived at a non-redundant list of 597 protein identifications with the potential for secretion either by classical secretory pathways or by non-secretory processes; 233 of them have been detected in cerebrospinal fluid or plasma as per the published literature, and 172 have been implicated in GBM or other cancers. The HNGC-2 secretome dataset could serve as a useful resource for designing a targeted investigation of GBM biomarkers in plasma.

Cx43-Associated Secretome and Interactome Reveal Synergistic Mechanisms for Glioma Migration and MMP3 Activation

“…SignalP (version 4.1, stringency 0.6) 4 (Petersen et al, 2011; Nielsen, 2017) observed 45 of these proteins having signals consistent with their production and export via the protein secretion pathway (Supplementary Table S1). Further inspection of our data using the Human Cancer Secretome Database (HCSD) 5 (Feizi et al, 2015) finds 98.8% (83/84) of C6/C6-13 proteins have been identified within the secretome of other cancers (Feizi et al, 2015); with 88% (74/84) observed with other GBM studies (Kashyap et al, 2010; Formolo et al, 2011; Gupta et al, 2013; Supplementary Table S1). Secretomes of both cell lines had strong functional links to Cellular Movement ( P = 2.15 × 10 -16 ), Cell Death and Survival (2.13 × 10 -15 ), Cancer (1.87 × 10 -12 ), and PTM (post-translational modification) (2.46 × 10 -11 ) (Figure 6A,C).…”

Proteomic Analysis on Anti-Proliferative and Apoptosis Effects of Curcumin Analog, 1,5-bis(4-Hydroxy-3-Methyoxyphenyl)-1,4-Pentadiene-3-One-Treated Human Glioblastoma and Neuroblastoma Cells

“…These interconnected glycolytic proteins catalyze the conversion of glucose to generate ATP, NADH, and pyruvate molecules to support the high load of anabolic reaction in cells. Overexpression of these proteins in the secretome of a glioblastoma cell line was associated with the progression of glioblastoma (Gupta et al, 2013). Li et al (2019) also showed that the growth and the progression of SH-SY5Y cells were significantly reduced following treatment with glycolysis inhibitory peptide.…”

Computational Screening of Secretory Proteins With Brain-Specific Expression in Glioblastoma Multiforme

“…SERPINI1 promote the mechanism for the initiation of brain metastasis in lung and breast cancers, by guarding tumor cells from death signals and promoting vascular cooption (Valiente et al 2014). Other members of this family are also involved in GBM pathogenesis; where SERPINE1 showed decreased expression in GBM cells (Tarassishin et al 2014), whereas SERPINA4, SERPINA10, SERPINC1 and SERPINF1 proteins exhibit signi cant downregulation in plasma of GBM patients (Gautam et al 2012;Gupta et al 2013). A strong interaction is evident for SERPINI1 with myc proto-oncogene protein (MYC) and protein max (Max), two prominent oncoproteins (Fig.…”