Evaluation of matrix metalloproteinase-2 (MMP-2) activity with film in situ zymography for improved cytological diagnosis of breast tumors.
BACKGROUND
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).


METHODS
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.


RESULTS
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.


CONCLUSIONS
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.
Fujiwara
Akira
A
Department of Pathology, Osaka University Graduate School of Medicine and Health Science, Japan. fujiwara_a@kich.itami.hyogo.jp.|Department of Molecular Pathology, Osaka University Graduate School of Medicine and Health Science, Japan. fujiwara_a@kich.itami.hyogo.jp.
Shibata
Eri
E
Department of Pharmacy, Osaka University Graduate School of Medicine and Health Science, Japan.
Terashima
Hitomi
H
Department of Pathology, Osaka University Graduate School of Medicine and Health Science, Japan.
Shishido
Akemi
A
Department of Pathology, Osaka University Graduate School of Medicine and Health Science, Japan.
Nishiki
Junko
J
Department of Pathology, Osaka University Graduate School of Medicine and Health Science, Japan.
Yoshida
Kyotaro
K
Department of Pathology, Osaka University Graduate School of Medicine and Health Science, Japan.
Miyauchi
Keisuke
K
Department of Surgery, Kinki Central Hospital, Osaka University Graduate School of Medicine and Health Science, Japan.
Madachi
Ayako
A
Department of Molecular Pathology, Osaka University Graduate School of Medicine and Health Science, Japan.
Matsuura
Nariaki
N
Department of Molecular Pathology, Osaka University Graduate School of Medicine and Health Science, Japan.
D002288:Adenocarcinoma, Mucinous / Q000175:diagnosis / Q000201:enzymology; D014408:Biomarkers, Tumor / Q000378:metabolism; D001943:Breast Neoplasms / Q000175:diagnosis / Q000201:enzymology*; D018270:Carcinoma, Ductal, Breast / Q000175:diagnosis / Q000201:enzymology; D018275:Carcinoma, Lobular / Q000175:diagnosis / Q000201:enzymology; D005260:Female; D005354:Fibrosarcoma / Q000175:diagnosis / Q000201:enzymology; D005780:Gelatin / Q000378:metabolism*; D006801:Humans; D007124:Immunoenzyme Techniques; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism*; D009361:Neoplasm Invasiveness / Q000473:pathology
D014408:Biomarkers, Tumor; C015233:gelatin film; D005780:Gelatin; D020778:Matrix Metalloproteinase 2