NPC2 as a Prognostic Biomarker for Glioblastoma Based on Integrated Bioinformatics Analysis and Cytological Experiments

Abstract

Glioblastoma (GBM) is one of the most common and fatal malignancies worldwide, while its prognostic biomarkers are still being explored. This study aims to identify potential genes with clinical and prognostic significance by integrating bioinformatics analysis and investigating their function in HNSCC. Based on the Single-cell RNA sequencing (scRNA-seq) results of H3K27M-glioma cells, computational bioinformatics methods were employed for selecting prognostic biomarker for GBM. The protein NPC2 (NPC Intracellular Cholesterol Transporter 2), which has been shown to be related to lipoprotein metabolism and innate immune system, was identified to be upregulated in GBM. NPC2 showed a relatively higher expression in GBM samples, and a negative correlation with tumor purity and tumor infiltrating immune cells. Additionally, NPC2 was knocked down in U87-MG and U251 cells line, and cell proliferation and migration capability were evaluated with CCK-8, scratch and transwell assay, respectively. Cytological experiments has shown that NPC2 overexpression inhibited GBM cells proliferation and migration, indicating its important role in GBM progression. This is the first investigation into the prognostic value of NPC2 interact with GBM. The potential molecular factor NPC2 have been identified as a prognostic biomarker for GBM.

Immune infiltration profiling in gastric cancer and their clinical implications

“…A large number of the genes in this model were closely related to the immune response. For example, NPC2 is involved in the lipoprotein metabolism and innate immune signal pathways,38…”

Cholesterol metabolism and its implication in glioblastoma therapy

“…LDL-derived cholesterol trapped in lysosomes imitates the absence of cholesterol uptake, thus inducing the inhibition of cell viability [68]. Since a study has demonstrated that NPC2 is an unfavorable prognostic biomarker in GBM [69], NPC2 may also become a potential target for the treatment of GBM. Ríos-Marco et al found that alkylphospholipids such as perifosine, edelfosine, erucylphosphocholine (ErPC) and hexadecylphosphocholine (HePC) interfere with cholesterol trafficking from the plasma membrane to the endoplasmic reticulum, hindering cholesterol esterification.…”

Hypomethylation-induced prognostic marker zinc finger DHHC-type palmitoyltransferase 12 contributes to glioblastoma progression

“…A CCK-8 assay was conducted as per our previous study (30). Briefly, cellular viability was determined using a CCK-8 (Beyotime Biotechnology).…”

“…A wound-healing assay was conducted as per our previous study (30). Briefly, DBTRG and U251 cells were seeded in a 6-well plate evenly with the cell concentration of 1×10 6 per well.…”

“…A transwell migration assay was conducted as per our previous study (30). Briefly, cells in the logarithmic growth phase were seeded in the upper transwell chamber insert (Corning, USA) at a density of 2×10 4 cells per well.…”