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Differential Expression of Matrix Metalloproteinases in Brain- and Bone-Seeking Clones of Metastatic MDA-MB-231 Breast Cancer Cells

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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 µM); epidermal growth factor and transforming growth factor β (20 ng/ml both). Migration and invasion assays were performed with the QCM™ 96-Well Migration/Invasion Assay (8 µm; Chemicon) over 24 h with or without specific MMPs inhibitors (MMP Inhibitor I Mix (5 µM); MMP-2/MMP-9 Inhibitor III (50 µM); 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.

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Acknowledgements

We thank Brigitte Rehmke, Bärbel Hufnagel and Jörg Krause for their expert technical assistance. This work was supported by the “Hensel-Foundation”.

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Authors and Affiliations

  1. Department of Neurosurgery, University of Kiel, Kiel, Germany

    Andreas M. Stark, Bartosz Anuszkiewicz, H. Maximilian Mehdorn & Janka Held-Feindt

  2. Institute of Anatomy, University of Kiel, Kiel, Germany

    Rolf Mentlein

  3. Division of Endocrinology and Metabolism, University of Texas Health Science Center, San Antonio, TX, USA

    Toshiyuki Yoneda

  4. Department of Neurosurgery, University of Schleswig-Holstein Medical Center, Campus Kiel, Schittenhelmstr. 10, 24105, Kiel, Germany

    Janka Held-Feindt

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  1. Andreas M. Stark
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Correspondence to Janka Held-Feindt.

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Stark, A.M., Anuszkiewicz, B., Mentlein, R. et al. Differential Expression of Matrix Metalloproteinases in Brain- and Bone-Seeking Clones of Metastatic MDA-MB-231 Breast Cancer Cells. J Neurooncol 81, 39–48 (2007). https://doi.org/10.1007/s11060-006-9207-0

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  • DOI: https://doi.org/10.1007/s11060-006-9207-0

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