Abstract
Emerging evidence suggests that the resistance of cancer stem cells (CSC) to many conventional therapies is one of the major limiting factors of cancer therapy efficacy. Identification of mechanisms responsible for survival and self-renewal of CSC will help design new therapeutic strategies that target and eliminate both differentiated cancer cells and CSC. Here we demonstrated the potential role of proapoptotic protein BAD in the biology of CSC in melanoma, prostate and breast cancers. We enriched CD44+/CD24− cells (CSC) by tumorosphere formation and purified this population by FACS. Both spheres and CSC exhibited increased potential for proliferation, migration, invasion, sphere formation, anchorage-independent growth, as well as upregulation of several stem cell-associated markers. We showed that the phosphorylation of BAD is essential for the survival of CSC. Conversely, ectopic expression of a phosphorylation-deficient mutant BAD induced apoptosis in CSC. This effect was enhanced by treatment with a BH3-mimetic, ABT-737. Both pharmacological agents that inhibit survival kinases and growth factors that are involved in drug resistance delivered their respective cytotoxic and protective effects by modulating the BAD phosphorylation in CSC. Furthermore, the frequency and self-renewal capacity of CSC was significantly reduced by knocking down the BAD expression. Consistent with our in vitro results, significant phosphorylation of BAD was found in CD44+ CSC of 83% breast tumor specimens. In addition, we also identified a positive correlation between BAD expression and disease stage in prostate cancer, suggesting a role of BAD in tumor advancement. Our studies unveil the role of BAD in the survival and self-renewal of CSC and propose BAD not only as an attractive target for cancer therapy but also as a marker of tumor progression.
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Abbreviations
- BAD:
-
BCL2-antagonist of cell death
- BrdU:
-
5-bromo-2′-deoxyuridine
- cdk:
-
cyclin-dependent kinase
- CSC:
-
cancer stem cell
- DC:
-
differentiated cancer
- FACS:
-
fluorescence-activated cell sorting
- E2:
-
17β-estradiol
- EGF:
-
epidermal growth factor
- MAPK:
-
mitogen-activated protein kinase
- PC:
-
parental cells
- PI:
-
propidium iodide
- shRNA:
-
small hairpin RNA
- TNFα:
-
tumor necrosis factor-alpha
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Acknowledgements
The authors thank Sara Tomei and Jingxuan Shan for help in statistical analyses and Aleksandra Liberska for FACS. This work was supported by the BMRP Funding of Weill Cornell Medical College in Qatar and Qatar Biomedical Research Institute.
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Sastry, K., Al-Muftah, M., Li, P. et al. Targeting proapoptotic protein BAD inhibits survival and self-renewal of cancer stem cells. Cell Death Differ 21, 1936–1949 (2014). https://doi.org/10.1038/cdd.2014.140
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DOI: https://doi.org/10.1038/cdd.2014.140
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