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Constitutive K-RasG12D activation of ERK2 specifically regulates 3D invasion of human pancreatic cancer cells via MMP-1.
- Source :
-
Molecular cancer research : MCR [Mol Cancer Res] 2012 Feb; Vol. 10 (2), pp. 183-96. Date of Electronic Publication: 2011 Dec 08. - Publication Year :
- 2012
-
Abstract
- Pancreatic ductal adenocarcinomas (PDAC) are highly invasive and metastatic neoplasms commonly unresponsive to current drug therapy. Overwhelmingly, PDAC harbors early constitutive, oncogenic mutations in K-Ras(G12D) that exist prior to invasion. Histologic and genetic analyses of human PDAC biopsies also exhibit increased expression of extracellular signal-regulated kinase (ERK) 1/2 and proinvasive matrix metalloproteinases (MMP), indicators of poor prognosis. However, the distinct molecular mechanisms necessary for K-Ras/ERK1/2 signaling and its influence on MMP-directed stromal invasion in primary human pancreatic ductal epithelial cells (PDEC) have yet to be elucidated in three-dimensions. Expression of oncogenic K-Ras(G12D) alone in genetically defined PDECs reveals increased invadopodia and epithelial-to-mesenchymal transition markers, but only when cultured in a three-dimensional model incorporating a basement membrane analog. Activation of ERK2, but not ERK1, also occurs only in K-Ras(G12D)-mutated PDECs cultured in three-dimensions and is a necessary intracellular signaling event for invasion based upon pharmacologic and short hairpin RNA (shRNA) inhibition. Increased active invasion of K-Ras(G12D) PDECs through the basement membrane model is associated with a specific microarray gene expression signature and induction of MMP endopeptidases. Specifically, MMP-1 RNA, its secreted protein, and its proteolytic cleavage activity are amplified in K-Ras(G12D) PDECs when assayed by real-time quantitative PCR, ELISA, and fluorescence resonance energy transfer (FRET). Importantly, shRNA silencing of MMP-1 mimics ERK2 inhibition and disrupts active, vertical PDEC invasion. ERK2 isoform and MMP-1 targeting are shown to be viable strategies to attenuate invasion of K-Ras(G12D)-mutated human pancreatic cancer cells in a three-dimensional tumor microenvironment.<br /> (©2011 AACR.)
- Subjects :
- Cells, Cultured
Enzyme Activation genetics
Epithelial-Mesenchymal Transition genetics
Gene Silencing
Humans
Matrix Metalloproteinase 1 metabolism
Mitogen-Activated Protein Kinase 1 metabolism
Mutation
Oligonucleotide Array Sequence Analysis
Pancreatic Ducts metabolism
Pancreatic Ducts pathology
Pancreatic Neoplasms metabolism
Pancreatic Neoplasms pathology
Signal Transduction
Gene Expression Regulation, Neoplastic
Genes, ras genetics
Matrix Metalloproteinase 1 genetics
Mitogen-Activated Protein Kinase 1 genetics
Neoplasm Invasiveness genetics
Subjects
Details
- Language :
- English
- ISSN :
- 1557-3125
- Volume :
- 10
- Issue :
- 2
- Database :
- MEDLINE
- Journal :
- Molecular cancer research : MCR
- Publication Type :
- Academic Journal
- Accession number :
- 22160930
- Full Text :
- https://doi.org/10.1158/1541-7786.MCR-11-0399