Human keratinocyte cell lines differ in the expression of the collagenolytic matrix metalloproteinases-1,-8, and-13 and of TIMP-1

Bachmeier BE, Nerlich AG, Boukamp P, Lichtinghagen R, Tschesche H, Fritz H, Fink E (2000)
BIOLOGICAL CHEMISTRY 381(5-6): 509-516.

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Bachmeier, BE; Nerlich, AG; Boukamp, P; Lichtinghagen, R; Tschesche, HaraldUniBi; Fritz, H; Fink, E
Abstract / Bemerkung
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.
MMP; collagenase; HaCaT cells; in vitro study; keratinocyte cell lines; tumorigenicity
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Bachmeier BE, Nerlich AG, Boukamp P, et al. Human keratinocyte cell lines differ in the expression of the collagenolytic matrix metalloproteinases-1,-8, and-13 and of TIMP-1. BIOLOGICAL CHEMISTRY. 2000;381(5-6):509-516.
Bachmeier, B. E., Nerlich, A. G., Boukamp, P., Lichtinghagen, R., Tschesche, H., Fritz, H., & Fink, E. (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.
Bachmeier, B. E., Nerlich, A. G., Boukamp, P., Lichtinghagen, R., Tschesche, H., Fritz, H., and Fink, E. (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, 509-516.
Bachmeier, B.E., et al., 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), p 509-516.
B.E. Bachmeier, et al., “Human keratinocyte cell lines differ in the expression of the collagenolytic matrix metalloproteinases-1,-8, and-13 and of TIMP-1”, BIOLOGICAL CHEMISTRY, vol. 381, 2000, pp. 509-516.
Bachmeier, B.E., Nerlich, A.G., Boukamp, P., Lichtinghagen, R., Tschesche, H., Fritz, H., Fink, E.: Human keratinocyte cell lines differ in the expression of the collagenolytic matrix metalloproteinases-1,-8, and-13 and of TIMP-1. BIOLOGICAL CHEMISTRY. 381, 509-516 (2000).
Bachmeier, BE, Nerlich, AG, Boukamp, P, Lichtinghagen, R, Tschesche, Harald, Fritz, H, and Fink, E. “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 (2000): 509-516.

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