Evaluation of matrix metalloproteinase-2 (mmp-2) activity with film in situ zymography for improved cytological diagnosis of Breast tumors

Abstract

Fine-needle aspiration (FNA) biopsy of breast tumors is a reliable diagnostic method for identifying breast carcinoma. However, it is sometimes difficult to definitely distinguish malignant from benign lesions. To improve the cytological diagnosis of breast tumors, we investigated the expression of active matrix metalloproteinase-2 (MMP-2), as detected by film in situ zymography (FIZ). We evaluated 34 fresh breast tumors, 25 paraffin-embedded breast tissue specimens, and a human cancer cell line (HT1080). MMP-2 expression was determined by immunocytochemistry or immunohistochemistry. Frozen sections and aspiration cytology samples of breast cancer were incubated on gelatin-coated films for the detection of active MMP-2. Immunohistochemistry showed that MMP-2 was expressed in cancer cells and stromal cells, but not in most benign breast lesions. Active MMP-2, but not pro-MMP-2, in the conditioned medium of HT1080 cells showed gelatinolytic activity on FIZ analysis, while the expression of both forms of MMP-2 was detected by immunocytochemistry. Gelatinolytic activity was also detected by FIZ analysis of aspiration cytology samples and frozen sections from the breast cancers, and there was a significant correlation between this gelatinolytic activity and the detection of MMP-2 expression by immunocytochemistry. The present study demonstrated that measurement of gelatinolytic activity by FIZ analysis of aspiration cytology samples may be useful for improving the cytological diagnosis of breast tumors.