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B.E. Bachmeier, A.G. Nerlich, P. Boukamp, R. Lichtinghagen, H. Tschesche, H. Fritz, E. Fink
Human Keratinocyte Cell Lines Differ in the Expression of the Collagenolytic Matrix Metallo-proteinases-1, -8, and -13 and of TIMP-1
We investigated cells and conditioned media of the three human keratinocyte cell lines HaCaT (nontumorigenic), 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 seminested RTPCR, Western blots, ELISA, immunocytochemistry and casein zymography. Transcripts of MMP-1, -8, -13 and TIMP-1 were detected in all cell lines by RTPCR 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 nontumorigenic 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.
Biological Chemistry, Walter de Gruyter
Print ISSN: 1431-6730
Volume: 381, 06/2000
Pages: 509 - 516