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PMID: 16166639 Published · ppublish English Journal Article Research Support, N.I.H., Extramural Research Support, U.S. Gov't, P.H.S.

The Runx2 osteogenic transcription factor regulates matrix metalloproteinase 9 in bone metastatic cancer cells and controls cell invasion.

Molecular and cellular biology ·Vol. 25 ·No. 19 ·2005-10-00 ·Pages 8581-91

Pratap J, Javed A, Languino LR, van Wijnen AJ, Stein JL, Stein GS, Lian JB

Abstract

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.

MeSH Terms
3T3 Cells Adenoviridae/genetics Animals Blotting, Western Bone Neoplasms/metabolism,pathology,secondary Cell Line, Tumor Cell Nucleus/metabolism Chromatin Immunoprecipitation Core Binding Factor Alpha 1 Subunit/physiology DNA, Complementary/metabolism Gene Expression Regulation, Enzymologic HeLa Cells Humans Matrix Metalloproteinase 9/biosynthesis,genetics Mice Mice, Mutant Strains Models, Biological Models, Genetic Neoplasm Invasiveness Neoplasm Metastasis Osteoblasts/metabolism Promoter Regions, Genetic RNA/chemistry RNA Interference RNA, Messenger/metabolism RNA, Small Interfering/metabolism Reverse Transcriptase Polymerase Chain Reaction Signal Transduction Transcriptional Activation
Chemicals
Core Binding Factor Alpha 1 Subunit DNA, Complementary RNA, Messenger RNA, Small Interfering RUNX2 protein, human RNA Matrix Metalloproteinase 9
Authors & Affiliations
7 authors, click to expand affiliations / ORCID
Pratap Jitesh
Department of Cell Biology, University of Massachusetts Medical School, 55 Lake Avenue North, Worcester, MA 01655, USA.
Javed Amjad
Languino Lucia R
van Wijnen Andre J
Stein Janet L
Stein Gary S
Lian Jane B
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Article Info
Journal
Molecular and cellular biology
Abbr.
Mol Cell Biol
ISSN
0270-7306
DOI
10.1128/MCB.25.19.8581-8591.2005
Published
2005-10-00
Pages
8581-91
Language
English
Region
United States
NLM ID
8109087
PMCID
PMC1265732
Subset
IM
Grants
NIAMS NIH HHS · P01 AR048818 · United States
NIAMS NIH HHS · R01 AR039588 · United States
NIDDK NIH HHS · P30 DK032520 · United States
NIAMS NIH HHS · P01 PO1 AR48818 · United States
NIDDK NIH HHS · P30 DK32520 · United States
NIAMS NIH HHS · AR39588 · United States
NIAMS NIH HHS · R01 AR049069 · United States
NCI NIH HHS · P01 CA082834 · United States
NCI NIH HHS · P01 CA82834 · United States
External Links
PubMed source DOI: 10.1128/MCB.25.19.8581-8591.2005 PMC Full Text View journal details
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