MMP-9-hemopexin domain hampers adhesion and migration of colorectal cancer cells.

Abstract

Matrix metalloproteinases (MMPs), in particular MMP-2 and MMP-9, are involved in colon cancer progression and metastasis due to their ability to degrade extracellular matrix (ECM) components. In previous studies we described the MMP-9 hemopexin like domain (MMP-9-PEX) as an MMP-9 antagonist. In the present study it was examined whether recombinant MMP-9-PEX has an inhibitory effect on migration and adhesion of colorectal carcinoma cells. Furthermore, we searched for MMP-9 substrate binding sites within the MMP-9-PEX by surface plasmon resonance. Migration of SW620 and LS174 cells was investigated in a modified Boyden chamber assay. In the presence of 0.2 microg/ml MMP-9-PEX migration of SW620 was decreased by 34%, while addition of 0.4 microg/ml diminished migration by 56%. Migration of LS174 cells was not affected by MMP-9-PEX. Adhesion studies were performed on 96-well plates coated with gelatin, collagen type I, and laminin, respectively. In the presence of MMP-9-PEX, adhesion of SW620 cells to these coating substrates was significantly inhibited. Surface plasmon resonance studies revealed binding of collagen type I and IV, elastin, and fibrinogen to proMMP-9 as well as to MMP-9-PEX. However, equilibrium constants (Kd) indicated a higher affinity of proMMP-9 to the matrix proteins. This could indicate that there is more than one binding site for matrix components within the entire proMMP-9 molecule. Since migration and adhesion of metastatic colorectal carcinoma cells were reduced by MMP-9-PEX, this recombinant MMP-9 antagonist might be of therapeutical interest.

• Authors: M Burg-Roderfeld, M Roderfeld, S Wagner, C Henkel, J Grötzinger, E Roeb
• Journal: International journal of oncology (Int J Oncol) Vol. 30 Issue 4 Pg. 985-92 (Apr 2007) ISSN: 1019-6439 [Print] Greece
• PMID: 17332939 (Publication Type: Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Antineoplastic Agents
• Collagen Type I
• Collagen Type IV
• Recombinant Fusion Proteins
• Recombinant Proteins
• Gelatin
• Fibrinogen
• Elastin
• Hemopexin
• MMP-9-hemopexin
• Matrix Metalloproteinase 9

Topics

• Antineoplastic Agents (isolation & purification, metabolism, pharmacology)
• Cell Adhesion (drug effects)
• Cell Line, Tumor
• Cell Movement (drug effects)
• Collagen Type I (metabolism)
• Collagen Type IV (metabolism)
• Colorectal Neoplasms (enzymology, pathology)
• Elastin (metabolism)
• Fibrinogen (metabolism)
• Gelatin (metabolism)
• Hemopexin (genetics, isolation & purification, metabolism, pharmacology)
• Humans
• Matrix Metalloproteinase 9 (genetics, isolation & purification, metabolism, pharmacology)
• Protein Structure, Tertiary (genetics)
• Recombinant Fusion Proteins
• Recombinant Proteins (isolation & purification, metabolism, pharmacology)
• Surface Plasmon Resonance