Proteomics identifies differentially expressed proteins in glioblastoma U87 cells treated with hederagenin.

IF 2.1 3区 生物学 Q3 BIOCHEMICAL RESEARCH METHODS Proteome Science Pub Date : 2023-04-29 DOI:10.1186/s12953-023-00208-7
Yesen Zhang, Yi Han, Yuchun Shang, Xiangyu Wang, Jiwei Sun
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Abstract

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.

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蛋白质组学鉴定了hederagenin处理的胶质母细胞瘤U87细胞中差异表达的蛋白质。
目的:研究hederagenin治疗人胶质母细胞瘤U87细胞后差异表达蛋白(DEPs)的表达情况,为hederagenin治疗胶质母细胞瘤提供理论依据。方法:采用细胞计数试剂盒8 (Cell Counting Kit 8)检测hederagenin对U87细胞增殖的抑制作用。通过串联质量标签和LC-MS/MS分析技术鉴定蛋白质。利用生物信息学方法对DEPs的注释、基因本体的富集和功能、京都基因基因组百科全书的通路和结构域进行了检测。根据TMT结果,从DEPs中选择hub蛋白进行WB验证。结果:蛋白定量分析共检出6522个蛋白。结论:Hederagenin对GBM U87细胞的抑制作用可能与KIF7有关,KIF7主要参与hedgehog信号通路。本研究结果为进一步研究hederagenin的治疗机制奠定了基础。
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来源期刊
Proteome Science
Proteome Science 生物-生化研究方法
CiteScore
2.90
自引率
0.00%
发文量
17
审稿时长
4.5 months
期刊介绍: Proteome Science is an open access journal publishing research in the area of systems studies. Proteome Science considers manuscripts based on all aspects of functional and structural proteomics, genomics, metabolomics, systems analysis and metabiome analysis. It encourages the submissions of studies that use large-scale or systems analysis of biomolecules in a cellular, organismal and/or environmental context. Studies that describe novel biological or clinical insights as well as methods-focused studies that describe novel methods for the large-scale study of any and all biomolecules in cells and tissues, such as mass spectrometry, protein and nucleic acid microarrays, genomics, next-generation sequencing and computational algorithms and methods are all within the scope of Proteome Science, as are electron topography, structural methods, proteogenomics, chemical proteomics, stem cell proteomics, organelle proteomics, plant and microbial proteomics. In spite of its name, Proteome Science considers all aspects of large-scale and systems studies because ultimately any mechanism that results in genomic and metabolomic changes will affect or be affected by the proteome. To reflect this intrinsic relationship of biological systems, Proteome Science will consider all such articles.
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