The Notch ligand Jagged2 promotes lung adenocarcinoma metastasis through a miR-200–dependent pathway in mice
Yanan Yang, … , Gregory J. Goodall, Jonathan M. Kurie
Yanan Yang, … , Gregory J. Goodall, Jonathan M. Kurie
Published March 14, 2011
Citation Information: J Clin Invest. 2011;121(4):1373-1385. https://doi.org/10.1172/JCI42579.
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Research Article Oncology

The Notch ligand Jagged2 promotes lung adenocarcinoma metastasis through a miR-200–dependent pathway in mice

Abstract

Epithelial tumor cells transit to a mesenchymal state in response to extracellular cues, in a process known as epithelial-to-mesenchymal transition (EMT). The precise nature of these cues has not been fully defined, an important issue given that EMT is an early event in tumor metastasis. Here, we have found that a population of metastasis-prone mouse lung adenocarcinoma cells expresses Notch and Notch ligands and that the Notch ligand Jagged2 promotes metastasis. Mechanistically, Jagged2 was found to promote metastasis by increasing the expression of GATA-binding (Gata) factors, which suppressed expression of the microRNA-200 (miR-200) family of microRNAs that target the transcriptional repressors that drive EMT and thereby induced EMT. Reciprocally, miR-200 inhibited expression of Gata3, which reversed EMT and abrogated metastasis, suggesting that Gata3 and miR-200 are mutually inhibitory and have opposing effects on EMT and metastasis. Consistent with this, high levels of Gata3 expression correlated with EMT in primary tumors from 2 cohorts of lung adenocarcinoma patients. These findings reveal what we believe to be a novel Jagged2/miR-200–dependent pathway that mediates lung adenocarcinoma EMT and metastasis in mice and may have implications for the treatment of human epithelial tumors.

Authors

Yanan Yang, Young-Ho Ahn, Don L. Gibbons, Yi Zang, Wei Lin, Nishan Thilaganathan, Cristina A. Alvarez, Daniel C. Moreira, Chad J. Creighton, Philip A. Gregory, Gregory J. Goodall, Jonathan M. Kurie

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Figure 7

miR-200 inhibits Gata3 expression.

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miR-200 inhibits Gata3 expression. (A) Forced miR-200 expression inhibit...
(A) Forced miR-200 expression inhibits Gata3. Quantitative PCR analysis of 344SQ cells subjected to forced miR-200b expression or empty lentiviral vector transfection (vector). Values represent the mean (± SD) of replicate (triplicate) samples. P values are from 2-tailed Welch’s t test. Q values estimate the fraction of comparisons with the given nominally significant P value that may arise from multiple testing. (B) Gata3 is not a direct miR-200 gene target. Reporter assays were performed using reporters fused to 3′-UTR sequences from Gata3 (RL-GATA3); ZEB1 (RL-Zeb1), which was included as a positive control; or nothing (RL-con). The Gata3 3′-UTR reporter construct (with positions of putative miR-200 binding sites) is illustrated graphically. These reporters were transiently transfected into miR-200b (miR200) or control 344SQ stable transfectants or were transiently cotransfected with synthetic miR-200 precursors (200a, 200b, or 205) or control oligomers (con) into 344SQ cells. Values were normalized based on renilla luciferase and expressed as the mean values (± SD) of replicate (triplicate) wells relative to those of controls cotransfected with empty reporter and empty expression vector or scrambled precursors, which were set at 1.0. Asterisks indicate F-test contrast P < 0.005 versus control. Values of 1-way ANOVA analysis are indicated.