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Thromb Haemost 2005; 93(04): 761-769
DOI: 10.1160/TH04-09-0601
Cellular Proteolysis and Oncology
Schattauer GmbH

Transcriptional control of cell density dependent regulation of matrix metalloproteinase and TIMP expression in breast cancer cell lines

Beatrice E. Bachmeier
1   Department of Clinical Chemistry and Clinical Biochemistry, Surgical Hospital, Ludwig-Maximilians-University Munich, Munich, Germany
,
Roberta Vené
2   Tumor Progression Unit, National Cance Research Institute, Genova, Italy
,
Cristina M. Iancu
1   Department of Clinical Chemistry and Clinical Biochemistry, Surgical Hospital, Ludwig-Maximilians-University Munich, Munich, Germany
3   Department of Biochemistry, University Bucharest, Romania
,
Ulrich Pfeffer
4   Functional Genomics, National Cancer Research Institute, Genova, Italy
,
Barbara Mayer
5   Department of Surgery Grosshadern Clinic, Ludwig-Maximilians-University Munich, Munich, Germany
,
Douglas Noonan
2   Tumor Progression Unit, National Cance Research Institute, Genova, Italy
,
Adriana Albini
6   Molecular Oncology, National Cancer Research Institute, Genova, Italy
,
Marianne Jochum
1   Department of Clinical Chemistry and Clinical Biochemistry, Surgical Hospital, Ludwig-Maximilians-University Munich, Munich, Germany
,
Andreas G. Nerlich
7   Institute of Pathology, Academic Hospital Munich-Bogenhausen, Munich, Germany
› Author AffiliationsFinancial support: Supported by DAAD/CRUI grant program Vigoni to B. B., and Associazione Italiana per la Ricerca sul Cancro (AIRC) and Ministero della Sanità to A. A.
Further Information

Publication History

Received 16 September 2004

Accepted after resubmission 16 March 2004

Publication Date:
14 December 2017 (online)

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Summary

Our recent studies on breast carcinoma cell lines with differing tumorigenicity / invasiveness (MCF-7< MDA-MB-468< MDAMB-231< MDA-MB-435) had shown significantly decreasing expression levels of MMPs-1,-2,-3,-8,-9,-10,-11 and –13 with increasing cell density while the levels of TIMP-1 and –2 increased. This correlated well with a lower invasiveness of confluent cells. In the present study, we extend our in vitro studies on three-dimensional cultures of breast cancer cell lines MCF-7 and MDAMB-435 and the transcriptional control of MMP and TIMP-expression in two-dimensional cultures of MDA-MB-231 and –435 cells. The tumor spheroid model showed that MMP expression and proteolytic activity were considerably higher in loosely structured tumor groups as compared to densely growing “compact” cell complexes. These data suggested that cell density regulates MMP and TIMP transcription and therefore, we tested whether AP-1, NFκB and CRE are involved in this process. Gene silencing of c-jun in sparse cultures had an inhibitory effect on MMP-3,–9 and –13 expression, on proteolytic activity as well as on the invasive potential of the cells, thus confirming a role for AP-1. TIMP-1, and –2 expression was up-regulated as compared to control cells. Consistent with this, overexpression of c-jun and c-fos in confluent breast cancer cell lines leads to up-regulation of MMP expression, proteolytic activity and invasion as well as down-regulation of TIMP-1. In summary, we provide evidence that cell density influences the invasive potential of tumor cells via regulation of MMPs and TIMPs by AP-1, NFκB and CRE transcription factors. Overexpression of MMPs in sparse cultures could help explain early dissemination of potentially metastatic cells.

Keywords

Breast cancer cell lines - MMP-regulation - gene silencing - invasion - cell density

Partly presented at the Second Chianti Meeting on Proteases held in Tuscany, Italy, from May 16–20, 2004


 

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