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Proteomics identifies differentially expressed proteins in glioblastoma U87 cells treated with hederagenin
Yesen Zhang1, Yi Han1, Yuchun Shang1
1Department of Neurosurgery, The First Affiliated Hospital of Bengbu Medical College, Bengbu, 233004, China.
Objective:
We investigated differentially expressed proteins (DEPs) in human glioblastoma U87 cells after treatment with hederagenin as a therapeutic screening mechanism and provided a theoretical basis for hederagenin in treating glioblastoma.
Methods:
The Cell Counting Kit 8 assay was used to analyze the inhibitory effect of hederagenin on the proliferation of U87 cells. Protein was identified by tandem mass tags and LC-MS/MS analysis techniques. Annotation of DEPs, Gene Ontology enrichment and function, and Kyoto Encyclopedia of Genes and Genomes pathways and domains were all examined by bioinformatics. According to the TMT results, hub protein was selected from DEPs for WB verification.
Results:
Protein quantitative analysis found 6522 proteins in total. Compared with the control group, 43 DEPs (P < 0.05) were involved in the highly enriched signaling pathway in the hederagenin group, among which 20 proteins were upregulated, and 23 proteins were downregulated. These different proteins are mainly involved in the longness regulating pathway-WORM, the hedgehog signaling pathway, Staphylococcus aureus infection, complement, coagulation cascades, and mineral absorption. KIF7 and ATAD2B expression were significantly down-regulated and PHEX and TIMM9 expression were significantly upregulated, according to WB analysis, supporting the TMT findings.
Conclusion:
Hederagenin inhibition of GBM U87 cells may be related to KIF7, which is mainly involved in the hedgehog signaling pathway. Our findings lay a foundation for additional study of the therapeutic mechanism of hederagenin.
Insights
Hederagenin effectively inhibits glioblastoma U87 cells by altering protein expression, particularly KIF7 within the hedgehog signaling pathway. This study provides a foundation for developing hederagenin as a glioblastoma treatment.
Area of Science:
- Molecular Biology
- Oncology
- Proteomics
Background:
- Glioblastoma (GBM) is an aggressive brain tumor with limited treatment options.
- Hederagenin, a natural compound, shows potential therapeutic effects.
- Understanding the molecular mechanisms of hederagenin in GBM is crucial for drug development.
Purpose of the Study:
- To investigate the differentially expressed proteins (DEPs) in human glioblastoma U87 cells treated with hederagenin.
- To identify the therapeutic potential of hederagenin for glioblastoma treatment.
- To provide a theoretical basis for hederagenin's application in glioblastoma therapy.
Main Methods:
- Cell Counting Kit 8 assay to assess hederagenin's effect on U87 cell proliferation.
- Tandem mass tags (TMT) and LC-MS/MS for protein identification and quantification.
- Bioinformatic analyses including Gene Ontology and Kyoto Encyclopedia of Genes and Genomes pathway enrichment.
- Western blot (WB) verification of selected hub proteins.
Main Results:
- Identification of 6522 proteins, with 43 differentially expressed proteins (DEPs) in hederagenin-treated U87 cells compared to controls.
- DEPs were significantly enriched in pathways including the hedgehog signaling pathway, complement, and coagulation cascades.
- Western blot confirmed downregulation of KIF7 and ATAD2B, and upregulation of PHEX and TIMM9, consistent with TMT findings.
Conclusions:
- Hederagenin's inhibition of glioblastoma U87 cells is potentially mediated by KIF7, a key component of the hedgehog signaling pathway.
- These findings support the investigation of hederagenin as a therapeutic agent for glioblastoma.
- Further research into the precise therapeutic mechanisms of hederagenin is warranted.
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