The Runx2 osteogenic transcription factor regulates matrix metalloproteinase 9 in bone metastatic cancer cells and controls cell invasion
Graduate School of Biomedical Sciences; Graduate School of Biomedical Sciences; Department of Cell Biology; Department of Cancer Biology
Medical Subject Headings
3T3 Cells; Adenoviridae; Animals; Blotting, Western; Bone Neoplasms; Cell Line, Tumor; Cell Nucleus; Chromatin Immunoprecipitation; Core Binding Factor Alpha 1 Subunit; DNA, Complementary; *Gene Expression Regulation, Enzymologic; Hela Cells; Humans; Matrix Metalloproteinase 9; Mice; Mice, Mutant Strains; Models, Biological; Models, Genetic; Neoplasm Invasiveness; Neoplasm Metastasis; Osteoblasts; Promoter Regions (Genetics); RNA; RNA Interference; RNA, Messenger; RNA, Small Interfering; Reverse Transcriptase Polymerase Chain Reaction; Signal Transduction; Trans-Activation (Genetics)
Life Sciences | Medicine and Health Sciences
The Runx2 (Cbfa1/AML3) transcription factor and matrix metalloproteinase 9 (MMP9) are key regulators of growth plate maturation and bone formation. The genes for both proteins are characteristic markers of breast and prostate cancer cells that metastasize to bone. Here we experimentally addressed the compelling question of whether Runx2 and MMP are functionally linked. By cDNA expression array analysis, we identified MMP9 as a novel downstream target of Runx2. Like that of MMP13, MMP9 expression is nearly depleted in Runx2 mutant mice. Chromatin immunoprecipitation and electrophoretic mobility shift assays revealed the recruitment of Runx2 to the MMP9 promoter. We show by mutational analysis that the Runx2 site mediates transactivation of the MMP9 promoter in osteoblasts (MC3T3-E1) and nonosseous (HeLa) cells. The overexpression of Runx2 by adenovirus delivery in nonmetastatic (MCF-7) and metastatic breast (MDA-MB-231) and prostate (PC3) cancer cell lines significantly increases the endogenous levels of MMP9. The knockdown of Runx2 by RNA interference decreases MMP9 expression, as well as that of other Runx2 target genes, including the genes for MMP13 and vascular endothelial growth factor. Importantly, we have demonstrated using a cell invasion assay that Runx2-regulated MMP9 levels are functionally related to the invasion properties of cancer cells. These results are consistent with Runx2 control of multiple genes that contribute to the metastatic properties of cancer cells and their activity in the bone microenvironment.
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Citation: Mol Cell Biol. 2005 Oct;25(19):8581-91. Link to article on publisher's site
Molecular and cellular biology
Pratap, Jitesh; Javed, Amjad; Languino, Lucia R.; Van Wijnen, Andre J.; Stein, Janet L.; Stein, Gary S.; and Lian, Jane B., "The Runx2 osteogenic transcription factor regulates matrix metalloproteinase 9 in bone metastatic cancer cells and controls cell invasion" (2005). GSBS Student Publications. 1003.