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- Title
Differential Expression of Matrix Metalloproteinases in Brain- and Bone-Seeking Clones of Metastatic MDA-MB-231 Breast Cancer Cells.
- Authors
Andreas Stark; Bartosz Anuszkiewicz; Rolf Mentlein; Toshiyuki Yoneda; H. Mehdorn
- Abstract
Abstract??Matrix Metalloproteinases (MMPs) play a crucial role in breast cancer metastasis. We examined the mRNA and protein expression of several MMPs in brain- and bone-seeking clones of MDA-MB-231 breast cancer cells, their transcriptional regulation and their functional role in the metastatic process. MMP mRNA expression was examined using real-time reverse transcription polymerase chain reaction. Protein expression was examined using enzyme linked immunosorbent essay (ELISA). The inducibility of mRNA and protein expression was tested with TPA (phorbol 12-myristate 13-acetate; 50??M); epidermal growth factor and transforming growth factor ? (20?ng/ml both). Migration and invasion assays were performed with the QCM? 96-Well Migration/Invasion Assay (8??m; Chemicon) over 24?h with or without specific MMPs inhibitors (MMP Inhibitor I Mix (5??M); MMP-2/MMP-9 Inhibitor III (50??M); EMD Biosciences). We found significantly higher mRNA expression of MMP-1 and -9 in brain-seeking 231-clones in comparison to -bone and -parental cells. In contrast, the mRNA expression of MMP-3 and -14 was comparable in all cells lines examined and MMP-13 expression was lower in both selective metastatic lines. MMP-2 and -8 were not expressed. ELISA revealed a higher amount of total as well as active MMP-1 and -9 in brain-seeking cells. TPA stimulation showed that MMP-1 and -9 transcription was inducible on the mRNA and protein level in 231-parental but not in 231-brain or -bone. 231-brain showed the highest migration and invasive capacity which could be decreased by the application of MMP-1 and/or MMP-9 inhibitor. Our results indicate functional importance of MMP-1 and -9 overexpression in brain metastasis in an in vitro model.
- Subjects
BREAST cancer; MESSENGER RNA; POLYMERASE chain reaction; METASTASIS
- Publication
Journal of Neuro-Oncology, 2007, Vol 81, Issue 1, p39
- ISSN
0167-594X
- Publication type
Article