Molecular cloning of canine membrane-anchored inhibitor of matrix metalloproteinase, RECK.
The reversion-inducing cysteine-rich protein with Kazal motifs (RECK) gene is one of the endogenous matrix metalloproteinase (MMP) inhibitors. It was reported that decreased RECK expression closely correlated with tumor malignancy. We determined the cDNA sequence of the canine RECK gene. The cDNA sequence and deduced amino acid of canine RECK were 2,913 bases and 971 residues, respectively. The predicted amino acid sequence of the protein showed 95.5% and 91.9% homology with human and mouse RECK, respectively. RECK mRNA expression was analyzed in various canine tissues and tumor cell lines by quantitative RT-PCR. The highest RECK expression was detected in lung and testis. In comparison with the tissues, a remarkably low expression level was detected in tumor cell lines. In addition, the RECK gene was transfected in the canine transitional cell carcinoma, and its influence on cell proliferation, migration, and invasion was analyzed. The transfected RECK gene suppressed only canine tumor invasion. These results showed that RECK might play an important role in tumor malignancy in dogs as well as in other mammalians.
Takagi
Satoshi
S
Laboratory of Veterinary Surgery, Department of Clinical Science, Graduate School of Veterinary Medicine, Hokkaido University, Sapporo, Japan.
Kitamura
Takanori
T
Hosaka
Yoshinao
Y
Ohsaki
Tomohiro
T
Bosnakovski
Darko
D
Kadosawa
Tsuyoshi
T
Okumura
Masahiro
M
Fujinaga
Toru
T
D000595:Amino Acid Sequence; D000818:Animals; D001483:Base Sequence; D045744:Cell Line, Tumor; D002465:Cell Movement / Q000235:genetics; D049109:Cell Proliferation; D003001:Cloning, Molecular; D017931:DNA Primers; D018076:DNA, Complementary / Q000235:genetics; D004285:Dogs / Q000235:genetics*; D058851:GPI-Linked Proteins; D015870:Gene Expression; D007150:Immunohistochemistry; D061965:Matrix Metalloproteinase Inhibitors; D008562:Membrane Glycoproteins / Q000235:genetics* / Q000378:metabolism; D008969:Molecular Sequence Data; D009361:Neoplasm Invasiveness / Q000235:genetics; D020133:Reverse Transcriptase Polymerase Chain Reaction; D016415:Sequence Alignment; D017422:Sequence Analysis, DNA; D017385:Sequence Homology; D014018:Tissue Distribution; D014162:Transfection
D017931:DNA Primers; D018076:DNA, Complementary; D058851:GPI-Linked Proteins; D061965:Matrix Metalloproteinase Inhibitors; D008562:Membrane Glycoproteins; C115460:RECK protein, human