紫外线照射后人晶状体上皮细胞的iTRAQ定量蛋白质组学研究
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国家自然科学基金面上项目(No.81974129,82171038,82101101)


Isobaric tags for relative and absolute quantitation quantitative proteomics research of human lens epithelial cells after ultraviolet-B irradiation
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National Natural Science Foundation of China(No.81974129, 82171038, 82101101)

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    摘要:

    目的:探索氧化损伤状态下人晶状体上皮细胞(SRA01/04)的蛋白表达变化,以期为年龄相关性白内障(ARC)发病机制的研究提供新的蛋白靶点。

    方法:将SRA01/04细胞分为实验组和对照组,实验组即细胞通过紫外线(UVB)照射以构建细胞氧化损伤模型,照射时长为10min; 对照组即细胞未经任何处理。应用同位素标记相对和绝对定量(iTRAQ)技术对这两组细胞进行蛋白组测序,以差异倍数>1.2且p<0.05的标准筛选差异表达蛋白(DEPs),并通过基因本体(GO)和京都基因和基因组百科全书(KEGG)数据库分别对上调和下调显著性前50的DEPs进行功能富集分析,利用Pathway commons构建蛋白-蛋白相互作用(PPI)网络。

    结果:本研究共筛选出552个DEPs。相比于对照组,实验组中上调的DEPs有176个,包括HMGB1、USP1等,下调的DEPs有376个,包括POLR2A、POLR2B等。GO和KEGG富集分析显示上调和下调显著性前50的DEPs参与了多种重要的生物学过程和信号通路。PPI网络提示氧化损伤修复(ODR)相关蛋白在UVB诱导的氧化损伤中可能起关键作用。

    结论:UVB照射SRA01/04细胞可致多种蛋白,尤其是ODR相关蛋白的表达发生改变,这些研究结果为深入探索ARC发病机制以及研究与治疗靶点相关的蛋白质或通路提供了细胞层面的参考。

    Abstract:

    AIM: To investigate the changes of protein expressions in human lens epithelial cells(SRA01/04)undergoing oxidative damage, hoping to provide new protein target for the pathogenesis of age-related cataract(ARC).

    METHODS: SRA01/04 cells were divided into experimental group and control group. In the experimental group, cells were irradiated with ultraviolet-B(UVB)for 10min to establish the model of oxidative damage, whereas cells in the control group were untreated. Protein expression profile from the two groups was sequenced by isobaric tags for relative and absolute quantitation(iTRAQ). The filtering criteria that fold change >1.2 and p<0.05 was used to determine the differentially expressed proteins(DEPs). Gene Ontology(GO)and Kyoto Encyclopedia of Genes and Genomes(KEGG)database were utilized for functional enrichment analysis of the top 50 DEPs with either up-regulated or down-regulated significance. Furthermore, Pathway commons software was used to establish the protein-protein interaction(PPI)network.

    RESULTS: Overall, 552 DEPs were screened out. A total of 176 DEPs were up-regulated in the experimental group compared with the control group, including HMGB1 and USP1, while 376 DEPs were down-regulated, including POLR2A and POLR2B. GO and KEGG enrichment analysis indicated that the top 50 DEPs with up-regulated or down-regulated significance were involved in various crucial biological processes and signaling pathways. PPI network revealed that oxidative damage repair(ODR)-related proteins might play a key role in UVB-induced oxidative damage.

    CONCLUSIONS: The expressions of multiple proteins, especially ODR-related proteins, can be altered in SRA01/04 cells via UVB irradiation. These findings may provide cellular-related insights into the pathogenesis of ARC and into proteins or pathways associated with therapeutic targets.

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陈晓娟,张国伟,李鹏飞,等.紫外线照射后人晶状体上皮细胞的iTRAQ定量蛋白质组学研究.国际眼科杂志, 2023,23(4):551-556.

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  • 收稿日期:2022-04-18
  • 最后修改日期:2023-03-07
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  • 在线发布日期: 2023-03-30
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