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.
View: Text | PDF
Research Article Oncology Article has an altmetric score of 3

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

×

Figure 4

Abrogation of TGF-β–induced EMT by depletion of Jagged2 but not Jagged1.

Options: View larger image (or click on image) Download as PowerPoint
Abrogation of TGF-β–induced EMT by depletion of Jagged2 but not Jagged1....
(A) Jagged depletion has no effect on Smad phosphorylation. Western blotting of total and phosphorylated (p) Smads in Jagged-depleted and control transfectants treated for the indicated time points with TGF-β. Actin indicates relative protein loading. (B) Loss of TGF-β–induced morphologic changes in Jagged2-depleted but not Jagged1-depleted or control 344SQ cells. Jagged2-depleted cells retain epithelial features after treatment for 48 hours with TGF-β (1 ng/ml) (bottom right panel). Original magnification, ×20. Scale bar: 100 μm. (C and D) Loss of TGF-β–induced EMT by Jagged2 depletion. Quantitative PCR analysis of (C) epithelial/mesenchymal markers and (D) transcriptional regulators of EMT in Jagged2-depleted and control shRNA transfectants treated for 48 hours with (black bars) or without (white bars) TGF-β (1 ng/ml). Values represent the mean (± SD) of replicate (triplicate) samples. P values are based on comparison of TGF-β–treated samples (control vs. Jagged2 depleted). Q values estimate the fraction of comparisons with the given nominally significant P value that may arise from multiple testing. (E) Western blotting of Snail1, Zeb1, Cdh2, Vim, and Actin in control and Jagged2-depleted cells treated with medium or TGF-β (1 ng/ml) for 48 hours. Actin indicates relative protein loading. Protein band density was quantified by densitometry, normalized to that of medium-treated control cells (which is set at 1.0), and is indicated below each blot.