miR-200–containing extracellular vesicles promote breast cancer cell metastasis
Minh T.N. Le, … , Leonora Balaj, Judy Lieberman
Minh T.N. Le, … , Leonora Balaj, Judy Lieberman
Published November 17, 2014
Citation Information: J Clin Invest. 2014;124(12):5109-5128. https://doi.org/10.1172/JCI75695.
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Research Article

miR-200–containing extracellular vesicles promote breast cancer cell metastasis

Abstract

Metastasis is associated with poor prognosis in breast cancer patients. Not all cancer cells within a tumor are capable of metastasizing. The microRNA-200 (miR-200) family, which regulates the mesenchymal-to-epithelial transition, is enriched in the serum of patients with metastatic cancers. Ectopic expression of miR-200 can confer metastatic ability to poorly metastatic tumor cells in some settings. Here, we investigated whether metastatic capability could be transferred between metastatic and nonmetastatic cancer cells via extracellular vesicles. miR-200 was secreted in extracellular vesicles from metastatic murine and human breast cancer cell lines, and miR-200 levels were increased in sera of mice bearing metastatic tumors. In culture, murine and human metastatic breast cancer cell extracellular vesicles transferred miR-200 microRNAs to nonmetastatic cells, altering gene expression and promoting mesenchymal-to-epithelial transition. In murine cancer and human xenograft models, miR-200–expressing tumors and extracellular vesicles from these tumors promoted metastasis of otherwise weakly metastatic cells either nearby or at distant sites and conferred to these cells the ability to colonize distant tissues in a miR-200–dependent manner. Together, our results demonstrate that metastatic capability can be transferred by the uptake of extracellular vesicles.

Authors

Minh T.N. Le, Peter Hamar, Changying Guo, Emre Basar, Ricardo Perdigão-Henriques, Leonora Balaj, Judy Lieberman

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Figure 2

miR-200 miRNAs are transferred from metastatic 4T1E cells to poorly metastatic 4TO7 cells.

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miR-200 miRNAs are transferred from metastatic 4T1E cells to poorly meta...
(A and B) miRNA and mRNA levels in 4T1E cells, untreated 4TO7 cells, and 4TO7-GFP cells sorted after 48-hour coculture with 4T1E cells in a 1:1 or 1:4 (4TO7/4T1E) ratio. miRNAs and mRNAs were quantified by TaqMan assay and SsoFast EvaGreen qRT-PCR, respectively (3 experiments). *P < 0.05; **P < 0.01, Student’s t test. (C) Western blot analysis in 4T1E and 4TO7-GFP cells that were untreated or cocultured with 4T1E cells for 1 week. (D) Flow cytometry analysis of E-cadherin (Ecad) staining in 4TO7-GFP cells that were untreated (U) or cocultured (C) with 4T1E cells at a 1:4 ratio for 1 week, gated on GFP+ single viable cells (3 experiments). *P < 0.05, Student’s t test. (E) E-cadherin staining (red) of 4T1E cells, 4TO7-GFP cells (green), and after 1 week, coculture of the 2 cell lines at a 1:4 ratio. DAPI (blue) marks nuclei. Scale bars: 10 μm.