Clin Orthop Surg.  2014 Dec;6(4):443-454. 10.4055/cios.2014.6.4.443.

Role of Matrix Metalloproteinase (MMP) 2 and MMP-9 in Soft Tissue Sarcoma

• Yang HK
• Jeong KC
• Kim YK
• Jung ST

• KMID: 1794734
• DOI: http://doi.org/10.4055/cios.2014.6.4.443

Abstract

BACKGROUND
We investigated the expression of matrix metalloproteinases (MMPs) and tissue inhibitors of metalloproteinases (TIMPs) in malignant fibrous histiocytoma (MFH), and determined whether these could be useful as prognostic factors.
METHODS
Among patients treated from 1993 to 2007, 30 cases of MFH were evaluated. Immunohistochemical staining was performed for MMP-2, MMP-9, TIMP-1, and TIMP-2 using paraffin wax-embedded blocks of MFH tissues. Reverse transcriptase polymerase chain reaction (RT-PCR) and Western blot and zymography were performed using fresh tissues obtained from 17 of the 30 cases. The levels of MMP and TIMP expression were compared between the MFH and normal control groups, and between non-metastatic and metastatic MFH groups.
RESULTS
Expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 were higher in the MFH group than the control group by RT-PCR, Western blotting, and zymography. Immunohistochemical staining revealed that MMP-2 and MMP-9 protein expression was higher in the metastatic than in the non-metastatic group. The expression levels of MMP-2 and TIMP-1 were significantly higher in the metastatic than in the non-metastatic group (p < 0.05) by RT-PCR. By Western blot analysis, the expression levels of MMP-2, TIMP-1, and TIMP-2 were higher in the metastatic group (p < 0.05), but MMP-9 showed only a slight increase in the metastatic group compared with the non-metastatic group (p > 0.05). Finally, gelatin zymography analysis showed that the expression levels of the pro- and active forms of MMP-2 were significantly higher in the metastatic group (p < 0.05), but the expression of the pro- and active forms of MMP-9 showed a slight decrease in the metastatic group (p > 0.05).
CONCLUSIONS
These results suggest that MMP-2, MMP-9, TIMP-1, and TIMP-2 may have important roles in the development and progression of MFH, and that the degree of expression of these metalloproteinases and their inhibitors, especially MMP-2, could be useful as prognostic factors related to metastasis in MFH.

Keyword

Matrix metalloproteinases; Tissue inhibitor of metalloproteinases; Malignant fibrous histiocytoma

MeSH Terms

Adult
Aged
Aged, 80 and over
Female
Histiocytoma, Malignant Fibrous/*metabolism
Humans
Immunohistochemistry
Male
Matrix Metalloproteinase 2/*biosynthesis
Matrix Metalloproteinase 9/*biosynthesis
Middle Aged
Neoplasm Metastasis
Prognosis
Tissue Inhibitor of Metalloproteinase-1/*biosynthesis
Tissue Inhibitor of Metalloproteinase-2/*biosynthesis
Matrix Metalloproteinase 2
Matrix Metalloproteinase 9
Tissue Inhibitor of Metalloproteinase-1
Tissue Inhibitor of Metalloproteinase-2

Figure

• 

  Fig. 1 Immunohistochemical staining findings for matrix metalloproteinase (MMP) 2, MMP-9, tissue inhibitors of metalloproteinase (TIMP) 1, and TIMP-2 in the non-metastatic malignant fibrous histiocytoma (MFH) and metastatic MFH cells. MMP-2 and MMP-9 were weakly expressed in non-metastatic MFH cells (A, C), but predominantly expressed in metastatic MFH cells with diffuse, strong intensities (B, D). TIMP-1 and TIMP-2 showed negative expressions in non-metastatic MFH cells (E, G) and focal strong expression in metastatic MFH cells (F, H) (A-H, ×200). (A) MMP-2 in non-metastatic MFH. (B) MMP-2 in metastatic MFH. (C) MMP-9 in non-metastatic MFH. (D) MMP-9 in metastatic MFH. (E) TIMP-1 in non-metastatic MFH. (F) TIMP-1 in metastatic MFH. (G) TIMP-2 in non-metastatic MFH. (H) TIMP-2 in metastatic MFH.

• 

  Fig. 2 The results of reverse transcriptase polymerase chain reaction for matrix metalloproteinase (MMP) 2, MMP-9, tissue inhibitors of metalloproteinase (TIMP) 1, and TIMP-2. The expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the non-metastatic malignant fibrous histiocytoma (MFH) and metastatic MFH groups were significantly higher than in the control group. The expression levels of MMP-2 and TIMP-1 were significantly higher and the expression levels of MMP-9 and TIMP-2 were lower in the metastatic MFH group than in the non-metastatic MFH group (p < 0.05). Meta: metastatic. *Significant, Mann-Whitney U-test, p < 0.05.

• 

  Fig. 3 Western blotting results for matrix metalloproteinase (MMP) 2, MMP-9, tissue inhibitors of metalloproteinase (TIMP) 1, and TIMP-2. The expression levels of MMP-2, MMP-9, TIMP-1, and TIMP-2 in the non-metastatic malignant fibrous histiocytoma (MFH) and metastatic MFH groups were higher than those in the control group, and these results were significant for MMP-9 and TIMP-2 (p < 0.05). The expression levels of MMP-2, TIMP-1, and TIMP-2 were higher in the metastatic MFH group than in the non-metastatic MFH group (p < 0.05). Meta: metastatic. *Significant, Mann-Whitney U-test, p < 0.05.

• 

  Fig. 4 Enzyme activity through gelatin zymography. Activities of the active forms of matrix metalloproteinase (MMP) 2 and MMP-9 were greater in the non-metastatic malignant fibrous histiocytoma (MFH) and metastatic MFH groups than in the control group. The expression levels of the pro- and active forms of MMP-2 were significantly higher and the expression levels of the pro- and active forms of MMP-9 decreased slightly in the metastatic MFH group than in the non-metastatic MFH group. Meta: metastatic. *Significant, Mann-Whitney U-test, p < 0.05.

• 

  Fig. 5 Results of reverse transcriptase polymerase chain reaction (A) and Western blotting (B) for matrix metalloproteinase (MMP) 14. Expression levels of MMP-14 in the control, non-metastatic malignant fibrous histiocytoma (MFH), and metastatic MFH groups were not statistically significantly different. Meta: metastatic. *Significant, Mann-Whitney U-test, p < 0.05.

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