MI-63: a novel small-molecule inhibitor targets MDM2 and induces apoptosis in embryonal and alveolar rhabdomyosarcoma cells with wild-type p53

Background: Interruption of the role of p53s as a tumour suppressor by MDM2 may be one of the mechanisms by which Cancer cells evade current therapy. Blocking the inhibition of wild-type p53 by MDM2 in Cancer cells should reactivate p53's tumour suppressor functions and enhance current Cancer treatments. MI-63 is a novel non-peptide small molecule that has shown strong binding affinity (K(i)=3 nM) for MDM2; however, its effects on paediatric Cancer cells and the specific mechanism of tumour suppressor reactivation have not been evaluated.

Methods: Rhabdomyosarcoma (RMS), the most common childhood soft tissue sarcoma, expresses either wild-type or mutant p53 protein. We examined the inhibitory effects of MI-63 in embryonal RMS (ERMS) and alveolar RMS (ARMS) cell lines expressing wild-type or mutated p53.

Results: Treatment with MI-63 reduced cell viability by 13.4% and by <1%, respectively, at 72 h in both RH36 and RH18 cell lines expressing wild-type p53. In contrast, RH30 and RD2 cells expressing p53 mutants are resistant to MI-63 treatment. An increased expression of p53, p21(WAF1), and Bax protein was observed after treatment with MI-63 in RMS cells with wild-type p53, and Apoptosis was confirmed by cleaved PARP and Caspase-3 expression. However, RD2 and RH30 RMS cells, as well as human normal skeletal muscle cells, showed a minimal increase in p53 signalling and no induction of cleaved PARP and Caspase-3. MI-63 was compared with Nutlin-3, a known MDM2 Inhibitor, and was found to be more potent in the inhibition of cell proliferation/viability. Further, synergy was observed when MI-63 was used in combination with doxorubicin.

Conclusion: These results indicate that MI-63 is a potent therapeutic agent for RMS cells expressing wild-type p53 protein.