MicroRNA-182 drives metastasis of primary sarcomas by targeting multiple genes
Mohit Sachdeva, Jeffrey K. Mito, Chang-Lung Lee, Minsi Zhang, Zhizhong Li, Rebecca D. Dodd, David Cason, Lixia Luo, Yan Ma, David Van Mater, Rebecca Gladdy, Dina C. Lev, Diana M. Cardona, David G. Kirsch
Mohit Sachdeva, Jeffrey K. Mito, Chang-Lung Lee, Minsi Zhang, Zhizhong Li, Rebecca D. Dodd, David Cason, Lixia Luo, Yan Ma, David Van Mater, Rebecca Gladdy, Dina C. Lev, Diana M. Cardona, David G. Kirsch
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Research Article

MicroRNA-182 drives metastasis of primary sarcomas by targeting multiple genes

Abstract

Metastasis causes most cancer deaths, but is incompletely understood. MicroRNAs can regulate metastasis, but it is not known whether a single miRNA can regulate metastasis in primary cancer models in vivo. We compared the expression of miRNAs in metastatic and nonmetastatic primary mouse sarcomas and found that microRNA-182 (miR-182) was markedly overexpressed in some tumors that metastasized to the lungs. By utilizing genetically engineered mice with either deletion of or overexpression of miR-182 in primary sarcomas, we discovered that deletion of miR-182 substantially decreased, while overexpression of miR-182 considerably increased, the rate of lung metastasis after amputation of the tumor-bearing limb. Additionally, deletion of miR-182 decreased circulating tumor cells (CTCs), while overexpression of miR-182 increased CTCs, suggesting that miR-182 regulates intravasation of cancer cells into the circulation. We identified 4 miR-182 targets that inhibit either the migration of tumor cells or the degradation of the extracellular matrix. Notably, restoration of any of these targets in isolation did not alter the metastatic potential of sarcoma cells injected orthotopically, but the simultaneous restoration of all 4 targets together substantially decreased the number of metastases. These results demonstrate that a single miRNA can regulate metastasis of primary tumors in vivo by coordinated regulation of multiple genes.

Authors

Mohit Sachdeva, Jeffrey K. Mito, Chang-Lung Lee, Minsi Zhang, Zhizhong Li, Rebecca D. Dodd, David Cason, Lixia Luo, Yan Ma, David Van Mater, Rebecca Gladdy, Dina C. Lev, Diana M. Cardona, David G. Kirsch

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Figure 2

Change in miR-182 levels modulates cell migration and invasion in primary mouse sarcoma cell lines.

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Change in miR-182 levels modulates cell migration and invasion in primar...
(A) Northern blot detects miR-182 in multiple primary sarcoma cell lines from KP mice. (B) Levels of miR-182 in cells correlates with the relative migration and invasion ability of these KP cells. Average of migration and invasion index was plotted on the y axis. (C) Real-time RT-PCR and (D) Northern blot validates knockdown of miR-182 in KP cell lines B and C by an anti–miR-182 oligo. (E) Knockdown of miR-182 in B and C decreases migration and (F) invasion (n = 6 independent experiments). (G) Stably transduced cell line A with anti–miR-182 lentiviral construct. (H) Northern blot validates stable knockdown of miR-182 in 2 different clones. (I and J) Stable knockdown of miR-182 does not affect orthotopic tumor growth in nude mice, but prevents lung metastasis after surgical resection of the orthotopic tumor, as shown in a Kaplan-Meier plot (K) (Mantel-Cox log-rank test). (L) Quantification of the number of lung metastases and (M) percentage of lung area with metastasis shows significant difference between the 2 genotypes. One-way ANOVA (C, E, and F) and 2-tailed Student’s t test (L and M) were used for statistical analysis. All data are mean ± SEM. Scale bars: 100 μm. *P < 0.05; **P < 0.01; ***P < 0.005.