HGF-MET signals via the MLL-ETS2 complex in hepatocellular carcinoma
Shugaku Takeda, … , Emily H. Cheng, James J. Hsieh
Shugaku Takeda, … , Emily H. Cheng, James J. Hsieh
Published June 24, 2013
Citation Information: J Clin Invest. 2013;123(7):3154-3165. https://doi.org/10.1172/JCI65566.
View: Text | PDF
Research Article

HGF-MET signals via the MLL-ETS2 complex in hepatocellular carcinoma

Abstract

HGF signals through its cognate receptor, MET, to orchestrate diverse biological processes, including cell proliferation, cell fate specification, organogenesis, and epithelial-mesenchymal transition. Mixed-lineage leukemia (MLL), an epigenetic regulator, plays critical roles in cell fate, stem cell, and cell cycle decisions. Here, we describe a role for MLL in the HGF-MET signaling pathway. We found a shared phenotype among Mll–/–, Hgf–/–, and Met–/– mice with common cranial nerve XII (CNXII) outgrowth and myoblast migration defects. Phenotypic analysis demonstrated that MLL was required for HGF-induced invasion and metastatic growth of hepatocellular carcinoma cell lines. HGF-MET signaling resulted in the accumulation of ETS2, which interacted with MLL to transactivate MMP1 and MMP3. ChIP assays demonstrated that activation of the HGF-MET pathway resulted in increased occupancy of the MLL-ETS2 complex on MMP1 and MMP3 promoters, where MLL trimethylated histone H3 lysine 4 (H3K4), activating transcription. Our results present an epigenetic link between MLL and the HGF-MET signaling pathway, which may suggest new strategies for therapeutic intervention.

Authors

Shugaku Takeda, Han Liu, Satoru Sasagawa, Yiyu Dong, Paul A. Trainor, Emily H. Cheng, James J. Hsieh

×

Figure 2

MLL functions downstream of the HGF-MET signaling pathway.

Options: View larger image (or click on image) Download as PowerPoint
MLL functions downstream of the HGF-MET signaling pathway. (A) Whole-mou...
(A) Whole-mount in situ hybridization analysis of E10.0 WT and Mll–/– embryos using specific RNA probes against Hgf and Met mRNA. No difference in Hgf and Met expression over the branchial arch region (white arrowhead) was noted. Representative images from 3 independent experiments are shown. Scale bars: 1 mm. (B) Quantitative RT-PCR analyses of Hgf and Met expression in tissues encompassing the hindbrain (orange outline) of WT and Mll–/– embryos. Data are mean ± SD. (C) MDCK scatter assay demonstrated equal competence of CM derived from WT and Mll–/– E10.5 primary MEFs. MDCK cells were allowed to form colonies for 24 hours and incubated in DMEM (Control) or the indicated CM for an additional 24 hours before imaging. (D) Ex vivo axon outgrowth assays. Bilateral hindbrain explants of rhombomeres 7 and 8 were cultured in collagen gel for 48 hours in the presence of embedded HGF-soaked and control mock-treated heparin acrylic beads. Explants from Mll–/– embryos showed impaired axon outgrowth toward HGF-soaked beads. Axons were immunostained using anti-neurofilament Ab, quantified in binary images using NIH ImageJ, and presented as the ratio of axon density toward HGF-treated versus control beads. Data are mean ± SD. *P < 0.05. Scale bars: 0.5 mm.