1. TIMP-1 increases expression and phosphorylation of proteins associated with drug resistance in breast cancer cells
- Author
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Maria Unni Rømer, Omid Hekmat, Irina Gromova, Louise Fogh, Lars Juhl Jensen, Anne-Sofie Schrohl, Jesper V. Olsen, Jan Stenvang, Stephanie Munk, Chiara Francavilla, Ramneek Gupta, Kirstine Belling, Ulrik Lademann, Niels Frank Jensen, Rachita Yadav, Sidse Ørnbjerg Würtz, Britt Damsgaard, Heiko Horn, Nils Brünner, Jose Moreira, and Dorte B. Bekker-Jensen
- Subjects
Proteome ,medicine.drug_class ,DNA repair ,Cell Survival ,Topoisomerase Inhibitors ,Molecular Sequence Data ,Gene Expression ,Antineoplastic Agents ,Breast Neoplasms ,Drug resistance ,Biology ,Biochemistry ,03 medical and health sciences ,0302 clinical medicine ,Stable isotope labeling by amino acids in cell culture ,Consensus Sequence ,medicine ,Humans ,Amino Acid Sequence ,Protein Interaction Maps ,Phosphorylation ,030304 developmental biology ,0303 health sciences ,Tissue Inhibitor of Metalloproteinase-1 ,Topoisomerase ,Phosphoproteomics ,General Chemistry ,Cell cycle ,3. Good health ,Cell biology ,DNA-Binding Proteins ,DNA Topoisomerases, Type II ,DNA Topoisomerases, Type I ,Drug Resistance, Neoplasm ,030220 oncology & carcinogenesis ,biology.protein ,MCF-7 Cells ,Female ,Cisplatin ,Protein Processing, Post-Translational ,Topoisomerase inhibitor - Abstract
Tissue inhibitor of metalloproteinase 1 (TIMP-1) is a protein with a potential biological role in drug resistance. To elucidate the unknown molecular mechanisms underlying the association between high TIMP-1 levels and increased chemotherapy resistance, we employed SILAC-based quantitative mass spectrometry to analyze global proteome and phosphoproteome differences of MCF-7 breast cancer cells expressing high or low levels of TIMP-1. In TIMP-1 high expressing cells, 312 proteins and 452 phosphorylation sites were up-regulated. Among these were the cancer drug targets topoisomerase 1, 2A, and 2B, which may explain the resistance phenotype to topoisomerase inhibitors that was observed in cells with high TIMP-1 levels. Pathway analysis showed an enrichment of proteins from functional categories such as apoptosis, cell cycle, DNA repair, transcription factors, drug targets and proteins associated with drug resistance or sensitivity, and drug transportation. The NetworKIN algorithm predicted the protein kinases CK2a, CDK1, PLK1, and ATM as likely candidates involved in the hyperphosphorylation of the topoisomerases. Up-regulation of protein and/or phosphorylation levels of topoisomerases in TIMP-1 high expressing cells may be part of the mechanisms by which TIMP-1 confers resistance to treatment with the widely used topoisomerase inhibitors in breast and colorectal cancer.
- Published
- 2013