Abstract
The mammalian target of rapamycin (mTOR) plays a central role in regulating the proliferation of malignant glioma cells, and mTOR-specific inhibitors such as rapamycin analogs are considered as promising therapy for malignant gliomas. However, the efficacy of mTOR inhibitors alone in the treatment of patients with malignant gliomas is only modest, potentially because these agents rather than acting as mTOR kinase inhibitors instead interfere with the function of only mTOR/raptor (regulatory-associated protein of mTOR) complex and thus do not perturb all mTOR functions. The purpose of this study was to determine whether global inhibition of the mTOR molecule enhances the antitumor effect of rapamycin on malignant glioma cells. We showed that rapamycin induced autophagy and that inhibition of autophagy by small interfering RNA (siRNA) directed against autophagy-related gene Beclin 1 attenuated the cytotoxicity of rapamycin in rapamycin-sensitive tumor cells, indicating that the autophagy was a primary mediator of rapamycin's antitumor effect rather than a protective response. Exogenous expression of an mTOR mutant interfering with its kinase activity markedly enhanced the incidence of rapamycin-induced autophagy. Moreover, silencing of mTOR with siRNA augmented the inhibitory effect of rapamycin on tumor cell viability by stimulating autophagy. Importantly, not only rapamycin-sensitive malignant glioma cells with PTEN mutations but also rapamycin-resistant malignant glioma cells with wild-type PTEN were sensitized to rapamycin by mTOR siRNA. These results indicate that rapamycin-induced autophagy is one of the agent's antitumor effects and that silencing or inhibiting mTOR kinase activity could enhance the effectiveness of rapamycin.
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Acknowledgements
We thank Dr Noboru Mizushima for the GFP-LC3 vector, Dr Robert T Abraham for the mTOR mutant SIDA vector and Emporia F Hollingsworth for technical assistance. We also thank Elizabeth L Hess for editing the manuscript. This work was supported from National Cancer Institute Grants CA088936 and CA108558, a start-up fund from The University of Texas MD Anderson Cancer Center (SK), a generous donation from the Anthony D. Bullock III Foundation (YK, SK) and Cancer Center Support Grant (CCSG)/Shared Resources of National Cancer Institute.
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Iwamaru, A., Kondo, Y., Iwado, E. et al. Silencing mammalian target of rapamycin signaling by small interfering RNA enhances rapamycin-induced autophagy in malignant glioma cells. Oncogene 26, 1840–1851 (2007). https://doi.org/10.1038/sj.onc.1209992
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DOI: https://doi.org/10.1038/sj.onc.1209992
