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The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44

Abstract

Cancer stem cells (CSCs), or tumor-initiating cells, are involved in tumor progression and metastasis1. MicroRNAs (miRNAs) regulate both normal stem cells and CSCs2,3,4,5, and dysregulation of miRNAs has been implicated in tumorigenesis6. CSCs in many tumors—including cancers of the breast7, pancreas8, head and neck9, colon10,11, small intestine12, liver13, stomach14, bladder15 and ovary16—have been identified using the adhesion molecule CD44, either individually or in combination with other marker(s). Prostate CSCs with enhanced clonogenic17 and tumor-initiating and metastatic18,19 capacities are enriched in the CD44+ cell population, but whether miRNAs regulate CD44+ prostate cancer cells and prostate cancer metastasis remains unclear. Here we show, through expression analysis, that miR-34a, a p53 target20,21,22,23,24, was underexpressed in CD44+ prostate cancer cells purified from xenograft and primary tumors. Enforced expression of miR-34a in bulk or purified CD44+ prostate cancer cells inhibited clonogenic expansion, tumor regeneration, and metastasis. In contrast, expression of miR-34a antagomirs in CD44 prostate cancer cells promoted tumor development and metastasis. Systemically delivered miR-34a inhibited prostate cancer metastasis and extended survival of tumor-bearing mice. We identified and validated CD44 as a direct and functional target of miR-34a and found that CD44 knockdown phenocopied miR-34a overexpression in inhibiting prostate cancer regeneration and metastasis. Our study shows that miR-34a is a key negative regulator of CD44+ prostate cancer cells and establishes a strong rationale for developing miR-34a as a novel therapeutic agent against prostate CSCs.

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Figure 1: Underexpression and tumor-inhibitory effects of miR-34a.
Figure 2: miR-34a inhibits prostate CSC properties.
Figure 3: Therapeutic effects of miR-34a.
Figure 4: CD44 is a direct and functional target of miR-34a.

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Acknowledgements

We thank K. Claypool and P. Whitney for FACS, the Histology Core for help with immunohistochemistry, K. Lin for statistical analysis, G. Calin for critically reading the manuscript and other members of the Tang lab for support and discussions. We also thank G. Hannon (Cold Spring Harbor Laboratory, Cold Spring Harbor, New York, USA) for the MSCV-PIG vector. This work was supported in part by grants from the US National Institutes of Health (R01-AG023374, R01-ES015888, R21-ES015893, R21-CA150009), the US Department of Defense (W81XWH-07-1-0616, W81XWH-08-1-0472) and Elsa Pardee Foundation (D.G.T.) and by two M.D. Anderson Cancer Center grants (CCSG-5 P30 CA016672-34 and ES007784). C. Liu and H. Li were supported in part by predoctoral fellowships from the US Department of Defense.

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Authors

Contributions

C.L., K.K., B.L., X.C. and L.P. designed and performed the experiments with help from C.J., T.C.-D., H.L., S.H., H.Y., J.F.W. and A.G.B., R.F. provided all HPCa samples. C.L. and D.G.T. prepared the manuscript. D.G.T., with help from D.B., designed the experiments and supervised the whole project. All authors discussed the results and commented on the manuscript.

Corresponding author

Correspondence to Dean G Tang.

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Competing interests

K.K, J.F.W, A.G.B. and D.B are employees of Mirna Therapeutics, Inc., which develops miRNA-based therapeutics.

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Supplementary Results, Supplementary Methods, Supplementary Figures 1–15 and Supplementary Tables 1 and 2 (PDF 2176 kb)

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Liu, C., Kelnar, K., Liu, B. et al. The microRNA miR-34a inhibits prostate cancer stem cells and metastasis by directly repressing CD44. Nat Med 17, 211–215 (2011). https://doi.org/10.1038/nm.2284

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