A rapid screening system evaluates novel inhibitors of DNA methylation and suggests F-box proteins as potential therapeutic targets for high-risk neuroblastoma
| Autor: | Christian Hagemeier, Livius Penter, Thomas Carell, Matthias Truss, Benjamin Hackner, Bert Maier, Ute Frede |
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| Rok vydání: | 2014 |
| Předmět: |
Cancer Research
DNA Methyltransferase Inhibitor Phthalimides Biology Pharmacology Chromatin remodeling Neuroblastoma Cell Line Tumor Antineoplastic Combined Chemotherapy Protocols medicine Humans Pharmacology (medical) Molecular Targeted Therapy Kinase activity Enzyme Inhibitors RNA Small Interfering DNA Modification Methylases Cell Proliferation Gene knockdown Cell growth F-Box Proteins Tryptophan Drug Synergism DNA Methylation medicine.disease DNA demethylation HEK293 Cells Pyrimidines Oncology Doxorubicin DNA methylation Cancer research Aminoquinolines Naphthoquinones |
| Zdroj: | Targeted oncology. 10(4) |
| ISSN: | 1776-260X |
| Popis: | After extensive research on radiochemotherapy, 5-year survival rates of children with high risk neuroblastoma still do not exceed 50%, owing to adverse side-effects exemplified by doxorubicin-induced cardiomyopathy. A promising new approach is the combination of conventional therapies with specific modulation of cell signaling pathways promoting therapeutic resistance, such as inhibition of aberrant kinase activity or re-expression of silenced tumor suppressor genes by means of chromatin remodeling. In this regard, we established a system that allows to identify potential drug targets as well as to validate respective candidate inhibitors in high-risk neuroblastoma model cell lines. Cell culture, drug exposure, shRNA-mediated knockdown and phenotype analysis are integrated into an efficient and versatile single well-based protocol. By utilizing this system, we assessed RG108, SGI-1027 and nanaomycin A, three novel DNA methyltransferase inhibitors that have not been tested in neuroblastoma cell lines so far, for their potential of synergistic anti-tumor activity in combination with doxorubicin. We found that, similarly to azacytidine, SGI-1027 and nanaomycin A mediate synergistic growth inhibition with doxorubicin independently of N-Myc status. However, they display high cytotoxicity but lack global DNA demethylation activity. Secondly, we conducted a lentiviral shRNA screen of F-box proteins, key regulators of protein stability, and identified Fbxw11/β-TrCP2 as well as Fbxo5/Emi1 as potential therapeutic targets in neuroblastoma. These results complement existing studies and underline the reliability and versatility of our single well-based protocol. |
| Databáze: | OpenAIRE |
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