1. The role of MDM2-NBS1 protein interaction in DNA repair in bladder cancer
- Author
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Aare, K and Kiltie, A
- Subjects
Bladder cancer ,DNA repair - Abstract
MDM2 is an E3 ubiquitin ligase that is frequently amplified in human cancers, including muscle-invasive bladder cancer. While it has a firmly established role in negative regulation of the tumour suppressor p53 by proteasomal degradation, the p53-independent functions of MDM2 remain scarcely understood. One of them is the delay of DNA double-strand break repair via a direct binding interaction with NBS1 of the MRE11-RAD50-NBS1 DNA repair complex. RG7388 is a selective MDM2 inhibitor, a second-generation member of the nutlin family that is currently in early clinical trials for solid and haematological tumours. The inhibition of MDM2-p53 protein interaction by RG7388 has no radiosensitising effect and does not disrupt the MDM2-NBS1 protein interaction in T24 and RT112 cells. However, compared to untreated controls, RG7388 alone and in combination with 5 Gy of ionising radiation (IR) alters the mRNA levels of MDM2 and NBS1 and protein levels of MDM2. NBS1 and MDM2 do not form any foci but co-localise in the nuclei of T24 and RT112 cells four hours after exposure to IR. In this study, we show that MDM2-NBS1 is an important protein interaction that remains intact in unstressed T24 and RT112 cells and following their exposure to genotoxic ionising radiation alone or in combination with RG7388. Under the same treatment conditions as well as in untreated controls, the pilot western blot experiment suggests that NBS1 is not ubiquitylated either in T24 or RT112 cells. Further experiments are required to verify this finding and to detect and characterise other post-translational modifications that may be involved in regulation of the MDM2-NBS1 protein interaction to determine its impact on DDR.
- Published
- 2020