Human Keratinocyte Cell Lines Differ in the Expression of the Collagenolytic Matrix Metalloproteinases-1, -8, and -13 and of TIMP-1

Beatrice E. Bachmeier, Hans Fritz, Edwin Fink, Andreas G. Nerlich, Petra Boukamp, Ralf Lichtinghagen, Harald Tschesche

    Research output: Contribution to journalArticle

    39 Scopus citations

    Abstract

    We investigated cells and conditioned media of the three human keratinocyte cell lines HaCaT (non-tumorigenic), A5 (benign, tumorigenic) and II-4RT (malignant, tumorigenic) with regard to production and secretion of the collagenases-1 to -3 (MMP-1, MMP-8 and MMP-13) and TIMP-1 using semi-nested RT-PCR, Western blots, ELISA, immunocytochemistry and casein zymography. Transcripts of MMP-1, -8, -13 and TIMP-1 were detected in all cell lines by RT-PCR and the corresponding proteins were found in the cytoplasm of all three cell lines by Western blot analysis and/or immunocytochemistry. The conditioned media of the malignant II-4RT cells contain significantly more MMP-1 and MMP-8 than those of HaCaT or A5 as evidenced by immunoblotting and ELISA. In addition to the presence of latent MMP-1, zymography also detected the active form of this enzyme. TIMP-1 was found only in extracts of all three cell lines, predominantly in A5. This study clearly indicates that the epithelial tumor cells synthesize different collagenases and TIMP-1. The malignant clone secretes increased amounts of distinct collagenases compared to the non-tumorigenic cell line, thereby verifying a correlation between biological behaviour and the amount of collagenases. In addition, we provide clear evidence that MMP-8 is not exclusively found in polymorphonuclear granulocytes, but also in keratinocyte cell lines.

    Original languageEnglish (US)
    Pages (from-to)509-516
    Number of pages8
    JournalBiological Chemistry
    Volume381
    Issue number5-6
    DOIs
    StatePublished - Jan 1 2000

    Keywords

    • Collagenase
    • HaCaT cells
    • In vitro study
    • Keratinocyte cell lines
    • MMP
    • Tumorigenicity

    ASJC Scopus subject areas

    • Biochemistry
    • Molecular Biology
    • Clinical Biochemistry

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  • Cite this

    Bachmeier, B. E., Fritz, H., Fink, E., Nerlich, A. G., Boukamp, P., Lichtinghagen, R., & Tschesche, H. (2000). Human Keratinocyte Cell Lines Differ in the Expression of the Collagenolytic Matrix Metalloproteinases-1, -8, and -13 and of TIMP-1. Biological Chemistry, 381(5-6), 509-516. https://doi.org/10.1515/BC.2000.065