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pmid:34188703

P4HA2 promotes cell proliferation and migration in glioblastoma.

Yuying Wu · Xunrui Zhang · Jue Wang · Ruijie Ji · Lei Zhang · Jianbing Qin · Meiling Tian · Guohua Jin · Xinhua Zhang

Oncology letters · 2021 doi:10.3892/ol.2021.12862

Paper

complete

Experiment

complete

Data

partial

Analysis

partial

Execution

absent

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Record details sci_version · id · kind · revision · content_hash · schema · created_at · updated_at · generator
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pmid:34188703
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title
P4HA2 promotes cell proliferation and migration in glioblastoma.
doi
10.3892/ol.2021.12862
pmid
34188703
authors array · 9 items
[0]
Yuying Wu
[1]
Xunrui Zhang
[2]
Jue Wang
[3]
Ruijie Ji
[4]
Lei Zhang
[5]
Jianbing Qin
[6]
Meiling Tian
[7]
Guohua Jin
[8]
Xinhua Zhang
journal
Oncology letters
year
2021
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.
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To validate the hub genes through survival analysis, the GEPIA2 website (http://gepia2.cancer-pku.cn/#index) was used, an online database for analysis of RNA expression levels, based on the Genotype-Tissue Expression Projects and The Cancer Genome Atlas (11). Kaplan-Meier survival curves were analyzed using the log-rank test.
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To validate the hub genes through survival analysis, the GEPIA2 website (http://gepia2.cancer-pku.cn/#index) was used, an online database for analysis of RNA expression levels, based on the Genotype-Tissue Expression Projects and The Cancer Genome Atlas (11). Kaplan-Meier survival curves were analyzed using the log-rank test.
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Basic characteristics of glioma stem cell and human glioblastoma
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To validate the hub genes through survival analysis, the GEPIA2 website (http://gepia2.cancer-pku.cn/#index) was used, an online database for analysis of RNA expression levels, based on the Genotype-Tissue Expression Projects and The Cancer Genome Atlas (11). Kaplan-Meier survival curves were analyzed using the log-rank test.
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HA2 overexpression and decreased after P4HA2-knockdown. In conclusion, the present study demonstrat
[1]
rials and methods Cell culture and transient transfection The U87 cell line (glioblastoma of unknown o
[2]
ed 5% CO2 incubator for 12 h, U87 cells were transfected with siRNAs (50 nM) using Lipofectamine® 300
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To validate the hub genes through survival analysis, the GEPIA2 website (http://gepia2.cancer-pku.cn/#index) was used, an online database for analysis of RNA expression levels, based on the Genotype-Tissue Expression Projects and The Cancer Genome Atlas (11). Kaplan-Meier survival curves were analyzed using the log-rank test.
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Complete file
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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. 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