Abstract
TIMP-1 (Tissue inhibitor of matrix metalloproteinase-1) is typically associated with inhibition of matrix metalloproteinases (MMP) induced invasion. However, TIMP-1 is overexpressed in many malignancies and is associated with poor prognosis in breast cancer. The mechanisms by which TIMP-1 promotes tumorigenesis are unclear. Reduced levels of TIMP-1 mediated by shRNA in MDA-MB-231 breast cancer cells had no effect on cellular physiology in vitro or tumor growth in SCID mice compared to vector control MDA-MB-231 cells. However, overexpression of TIMP-1 in MDA-MB-231 cells resulted in inhibition of cell invasion and enhanced phosphorylation of p38 MAPK and AKT in vitro. Additionally, treatment of parental MDA-MB-231 cells with purified TIMP-1 protein led to activation of p38 MAPK and MKK 3/6. cDNA array analysis demonstrated that high expression of TIMP-1 in MDA-MB-231 cells resulted in alterations in expression of approximately 200 genes, 1.5 fold or greater compared to vector control cells (P < 0.1). Real-time RT-PCR confirmed changes in expression of several genes associated with cancer progression including DAPK1, FGFR4 and MAPK13. In vivo, high TIMP-1 expression induced tumor growth in SCID mice compared to vector control cells and increased tumor vessel density. Affymetrix array analysis of vector control and TIMP-1 MDA-MB-231 xenograft tumors revealed that TIMP-1 altered expression of approximately 600 genes in vivo, including MMP1, MMP13, S100A14, S100P, Rab25 and ID4. These combined observations suggest that the effects of TIMP-1 differ significantly in a 2-D environment compared to the 3-D environment and that TIMP-1 stimulates tumor growth.
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The work was funded through the Department of Defense, Grant # BC020675 (JAC) and the National Institute of Health, NRSA, Fellowship #CA112714 (RLHB).
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Supplementary Table 1
Affymetrix cDNA array analysis of MDA-MB-231 pcDNA pool #1 and TIMP-1 pool #1 breast cancer cells in monolayer culture (XLS 51 kb)
Supplementary Table 2
Affymetrix cDNA array analysis of MDA-MB-231 pcDNA pool #1 and TIMP-1 pool #1 MDA-MB-231 breast cancer xenografts (XLS 148 kb)
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Bigelow, R.L.H., Williams, B.J., Carroll, J.L. et al. TIMP-1 overexpression promotes tumorigenesis of MDA-MB-231 breast cancer cells and alters expression of a subset of cancer promoting genes in vivo distinct from those observed in vitro. Breast Cancer Res Treat 117, 31–44 (2009). https://doi.org/10.1007/s10549-008-0170-7
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DOI: https://doi.org/10.1007/s10549-008-0170-7