Quantitative Proteomics Analysis Reveals Nuclear Perturbation in Human Glioma U87 Cells treated with Temozolomide
Quantitative Proteomics Analysis Reveals Nuclear Perturbation in Human Glioma U87 Cells treated with Temozolomide
复制标题
定量蛋白质组学分析揭示用替莫唑胺处理的人胶质瘤 U87 细胞的核扰动
DOI：
10.1002/cbf.3459
复制
发表时间：
2019-12-12
影响因子：
3.6
通讯作者：
Liu, Yawei
中科院分区：
文献类型：
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作者：
Guo, Jinglin;Yi, Guo-zhong;Liu, Yawei
胶质母细胞瘤（GBM）是恶性程度最高、侵袭性最强的胶质瘤，预后极差。替莫唑胺（TMZ）仍然是一线治疗，但即使在MGMT缺陷的胶质母细胞瘤细胞中，耐药性也是不可避免的。本研究的目的是了解TMZ对细胞核的影响以及MGMT缺陷GBM获得性TMZ耐药的机制。我们显示了TMZ处理1周后MGMT缺陷的GBM U87细胞核蛋白质组的变化。采用基于无标记的定量蛋白质组学研究核蛋白丰度变化。随后，基因本体功能注释，KEGG通路分析，蛋白质-蛋白质相互作用（PPI）网络构建分析的DAP，和免疫荧光来验证蛋白质组学的质量。总共鉴定了457个（455个基因产物）显著DAP，其中327个上调，128个下调。生物信息学分析发现，RAD 50、MRE 11、UBR 5、MSH 2、MSH 6、DDB 1、DDB 2、RPA 1、RBX 1、CUL 4A和CUL 4 B主要富集DNA损伤修复相关通路，构成蛋白质-蛋白质相互作用网络。核糖体蛋白表达下调。细胞处于应激反应状态，而整个代谢水平降低。TMZ处理U87细胞1周后，细胞核内多种蛋白质表达异常，导致DNA损伤。一些与DNA损伤修复途径相关的蛋白表达上调，且存在较强的相互作用。我们认为这是肿瘤细胞化疗耐药性的潜在线索。这些蛋白质可作为未来肿瘤耐药性筛选的指标。
Glioblastoma (GBM) is the most malignant and aggressive glioma, which has a very poor prognosis. Temozolomide (TMZ) is still a first-line treatment, but resistance is inevitable even in MGMT-deficient glioblastoma cells. The aims of this study were to comprehend the effect of TMZ on nucleus and the underlying mechanism of acquired TMZ resistance in MGMT-deficient GBM. We show the changes of nuclear proteome in the MGMT-deficient GBM U87 cells treated with TMZ for 1 week. Label-free-based quantitative proteomics were used to investigate nuclear protein abundance change. Subsequently, gene ontology function annotation, KEGG pathway analysis, protein-protein interaction (PPI) network construction analysis of DAPs, and immunofluorescence were applied to validate the quality of proteomics. In total, 457 (455 gene products) significant DAPs were identified, of which 327 were up-regulated and 128 were down-regulated. Bioinformatics analysis uncovered RAD50, MRE11, UBR5, MSH2, MSH6, DDB1, DDB2, RPA1, RBX1, CUL4A, and CUL4B mainly enriched in DNA damage repair related pathway and constituted a protein-protein interaction network. Ribosomal proteins were down-regulated. Cells were in a stress-responsive state, while the entire metabolic level was lowered. Significance of the study In U87 cell treated with TMZ for 1 week, which resulted in DNA damage, we found various proteins dysregulated in the nucleus. Some proteins related to the DNA damage repair pathway were up-regulated, and there was a strong interaction. We believe this is the potential clues of chemotherapy resistance in tumour cells. These proteins can be used as indicators of tumour resistance screening in the future.