P4HA2 promotes cell proliferation and migration in glioblastoma

Abstract

Glioblastoma (GBM) is a primary malignant tumor characterized by high infiltration and angiogenesis in the brain parenchyma. Glioma stem cells (GSCs), a heterogeneous GBM cell type with the potential for self-renewal and differentiation to tumor cells, are responsible for the high malignancy of GBM. The purpose of the present study was to investigate the roles of significantly differentially expressed genes between GSCs and GBM cells in GBM progression. The gene profiles GSE74304 and GSE124145, containing 10 GSC samples and 12 GBM samples in total, were obtained from the Gene Expression Omnibus (GEO) database. The overlapping differentially expressed genes were identified with GEO2R tools and Venn software online. Subsequently, Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway analysis was performed on the 41 upregulated and 142 downregulated differentially expressed genes in GSCs compared with in GBM cells via the DAVID website. Protein-protein interaction and module analyses in Cytoscape with the STRING database revealed 21 hub genes that were downregulated in GSCs compared with in GBM cells. Survival analysis conducted via the GEPIA2 website revealed that low expression levels of the hub genes prolyl 4-hydroxylase subunit α2 (P4HA2), TGF-β induced, integrin subunit α3 and thrombospondin 1 were associated with significantly prolonged survival time in patients with GBM. Further experiments were performed focusing on P4HA2. Reverse transcription-quantitative PCR was used to detect P4HA2 gene expression. In agreement with the bioinformatics analysis, P4HA2 expression was higher in U87 cells than in GSCs. Cell Counting Kit-8, EdU incorporation, cell cycle analysis, wound healing and Transwell assays demonstrated that the cell proliferation and migration increased after P4HA2 overexpression and decreased after P4HA2-knockdown. In conclusion, the present study demonstrated that low P4HA2 expression in GSCs promoted GBM cell proliferation and migration, suggesting that P4HA2 may act as a switch in the transition from GSCs to GBM cells.

CEBPB upregulates P4HA2 to promote the malignant biological behavior in IDH1 wildtype glioma

“…9 Consequently, collagen deposition interferes with cell polarity and adhesion, ultimately promoting cancer cell proliferation 34 and facilitating cancer cell invasion. 35 Previous studies found that P4HA2 was upregulated in GBM and promoted the proliferation, migration, and invasion of glioma cells, 36,37 consistent with our study. Unlike these previous studies, we found an unusual transcriptional expression pattern of P4HA2 that was at the lowest in LGG, in comparison to normal brain tissues and HGG.…”

COL5A1 Serves as a Biomarker of Tumor Progression and Poor Prognosis and May Be a Potential Therapeutic Target in Gliomas

“…Furthermore, the PPI networks of the co-expressed genes suggested that collagen members, such as COL8A1, COL1A1, COL1A2, and the like, were the main connected components with COL5A1. Prolyl-4-hydroxylase subunit 2 (P4HA2), as one of the hub genes interacted the collagen members above, was reported to play an important role in tumor proliferation, migration, invasion, and epithelial-to-mesenchymal transition (EMT) in gliomas ( 32 , 33 ). Another similar hub gene Serpin family H member 1 (SERPINH1), also known as HSP47, was proved to promote GBM stem-like cell survival by modulating tumor microenvironment through TGF-β pathway ( 34 ) and enhance glioma angiogenesis through HIF1α-VEGFR2 signaling ( 35 ).…”

Lama2 And Samsn1 Mediate the Effects of Brn4 on Hippocampal Neural Stem Cell Proliferation and Differentiation

“…For primary hippocampal NSCs, the RNA was extracted 7 days after induced differentiation in DMEM/F-12 supplementing with 1% FBS and 100 U/mL penicillin-streptomycin 24 h after plasmid transfection or 48 h after lentivirus infection at 37°C with 5% CO 2 . As described previously [ 26 ], total RNA was extracted using Total RNA Isolation Reagent (Shanghai Pufei Biotechnology), cDNA synthesis was performed with a RevertAid First Strand cDNA Synthesis kit (Thermo Fisher Scientific), and RT-qPCR was performed with Universal SYBR Green Master Mix (Roche Diagnostics) according to the manufacturer's protocol using the following PCR conditions: 95°C for 10 min, followed by 40 cycles of 95°C for 30 s, 60°C for 30 s and 72°C for 40 s, followed by a final elongation step of 10 min at 72°C. Gene expression was normalized to β -actin expression calculated using the 2 −ΔΔCT method [ 27 ].…”

“…Cell Cycle Analysis. As described [26], NSCs were harvested 24 h after plasmid transfection or 48 h after lentivirus infection and then fixed with ice-cold 75% ethanol at -20 °C for at least 24 h. Next, cells were stained with Hoechst 33342 solution (BD Biosciences) and incubated for 30 min at RT, washed, and suspended in BD Pharmingen™ Stain Buffer (BD Biosciences). The results were analyzed using the BD FACSCalibur system (BD Biosciences) with ModFit LT v3.3.11 software (Verity Software House).…”