Abstract
Murine double minute (MDM2) binding protein (MTBP) has been implicated in cancer progression. Here, we demonstrate one mechanism by which MTBP inhibits cancer metastasis. Overexpression of MTBP in human osteosarcoma cell lines lacking wild-type p53 did not alter primary tumor growth in mice, but significantly inhibited metastases. MTBP downregulation increased the migratory potential of MDM2−/−p53−/− mouse embryonic fibroblasts, suggesting that MTBP inhibited cell migration independently of the Mdm2–p53 pathway. Co-immunoprecipitation and mass spectrometric analysis identified alpha-actinin-4 (ACTN4) as an MTBP-interacting protein. Endogenous MTBP interacted with and partially colocalized with ACTN4. MTBP overexpression inhibited cell migration and filopodia formation mediated by ACTN4. Increased cell migration by MTBP downregulation was inhibited by concomitant downregulation of ACTN4. MTBP also inhibited ACTN4-mediated F-actin bundling. We furthermore demonstrated that nuclear localization of MTBP was dispensable for inhibiting ACTN4-mediated cell migration and filopodia formation. Thus, MTBP suppresses cell migration, at least partially, by inhibiting ACTN4 function. Our study not only provides a mechanism of metastasis suppression by MTBP, but also suggests MTBP as a potential biomarker for cancer progression.
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Abbreviations
- ACTN4:
-
alpha-actinin-4
- DMSO:
-
dimethyl sulfoxide
- DAPI, 4′,6-diamidino-2-phenylindole; MDM2:
-
murine double minute
- MTBP:
-
MDM2 binding protein
- MEFs:
-
mouse embryonic fibroblasts
- PBS:
-
phosphate buffered saline
- siRNA:
-
short interfering RNA
- shRNA:
-
short hairpin RNA
- SDS:
-
sodium dodecyl sulfate.
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Acknowledgements
We thank Drs Indu Kheterpal, Yuki Tochigi and Louis Marrero for their technical assistance. We also thank Ms Kristy-Le T Nguyen for her technical assistance and editing the manuscript. This work was supported by grants from P20 RR020152-02 (PLD), RSG-09-169-01-CSM (JI), LCRC start-up (TI) and National Foundation for Cancer Research-Center for Metastasis Research (DRW).
Author contribution: Neeraj Agarwal, Amit S Adhikari, Swathi V Iyer, Kevon Hekmatdoost and Tomoo Iwakuma all performed the experiments. Danny R Welch provided us with the crucial materials and discussion. Neeraj Agarwal, Swathi V Iyer and Tomoo Iwakuma wrote the manuscript.
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Agarwal, N., Adhikari, A., Iyer, S. et al. MTBP suppresses cell migration and filopodia formation by inhibiting ACTN4. Oncogene 32, 462–470 (2013). https://doi.org/10.1038/onc.2012.69
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DOI: https://doi.org/10.1038/onc.2012.69
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