High expression of 92 kDa type IV collagenase (matrix metalloproteinase-9) in canine mammary adenocarcinoma.
The 92 kDa type VI collagenase (matrix metalloproteinase-9 (MMP-9)) activities on zymography assay were found to be 1-6 times higher in benign tumor breast tissues of 12 canines and 4-26 times higher in adenocarcinoma breast tissues of nine canines than that of control tissues, respectively. A full-length canine MMP-9 cDNA was cloned from the adenocarcinoma tissue by reverse transcription-PCR and 5'- and 3'-RACE. The isolated cDNA contained an open reading frame coding for a polypeptide of 704 amino acids. The predicted protein sequence displayed extensive similarity to that of known MMP-9s and contained a putative signal sequence, a propeptide, an active site with three zinc-binding histidine residues, a calcium-binding domain, a hemopexin region, and three key cysteine residues. Western blotting using MMP-9-specific antibodies prepared against the peptide corresponding to Arg(642)-Asp(704) of canine MMP-9 and Northern blotting using a MMP-9-specific cDNA fragment as a probe confirmed that MMP-9 (the 92 kDa protein band) was highly expressed in canine mammary adenocarcinoma tissues. Higher levels of MMP-9 activity were found in the sera of canines with mammary adenocarcinoma. The results indicated that MMP-9 plays an important role in the progression of a canine mammary tumor and that assay of serum MMP-9 is helpful for early diagnosis as progress of adenocarcinoma.
Yokota
H
H
Department of Veterinary Biochemistry, School of Veterinary Medicine, Rakuno Gakuen University, Ebetsu, Hokkaido, Japan. h-yokota@rakuno.ac.jp
Kumata
T
T
Taketaba
S
S
Kobayashi
T
T
Moue
H
H
Taniyama
H
H
Hirayama
K
K
Kagawa
Y
Y
Itoh
N
N
Fujita
O
O
Nakade
T
T
Yuasa
A
A
D000230:Adenocarcinoma / Q000097:blood / Q000201:enzymology* / Q000662:veterinary; D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D014408:Biomarkers, Tumor / Q000097:blood; D003001:Cloning, Molecular; D018076:DNA, Complementary / Q000096:biosynthesis / Q000737:chemistry; D004283:Dog Diseases / Q000097:blood / Q000201:enzymology*; D004285:Dogs; D015674:Mammary Neoplasms, Animal / Q000097:blood / Q000201:enzymology*; D020780:Matrix Metalloproteinase 9 / Q000096:biosynthesis* / Q000097:blood / Q000737:chemistry / Q000235:genetics; D008969:Molecular Sequence Data; D011379:Prognosis
D014408:Biomarkers, Tumor; D018076:DNA, Complementary; D020780:Matrix Metalloproteinase 9