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Quantitative Proteomics Analysis Reveals Nuclear Perturbation in Human Glioma U87 Cells treated with Temozolomide
Author(s) -
Guo Jinglin,
Yi Guozhong,
Liu Zhifeng,
Sun Xuegang,
Yang Runwei,
Guo Manlan,
Li Yaomin,
Li Ke,
Li Kaishu,
Wang Xiran,
Song Haimin,
Qi Songtao,
Huang Guanglong,
Liu Yawei
Publication year - 2020
Publication title -
cell biochemistry and function
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 0.933
H-Index - 61
eISSN - 1099-0844
pISSN - 0263-6484
DOI - 10.1002/cbf.3459
Subject(s) - temozolomide , proteomics , kegg , dna repair , msh6 , stable isotope labeling by amino acids in cell culture , glioma , cancer research , u87 , dna damage , rad50 , nuclear protein , biology , mg132 , quantitative proteomics , proteome , microbiology and biotechnology , bioinformatics , gene , proteasome inhibitor , proteasome , dna binding protein , dna , biochemistry , gene expression , transcriptome , dna mismatch repair , transcription factor
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.