Membrane-type 1 matrix metalloproteinase enhances lymph node metastasis of gastric cancer.
The mechanisms of the lymph node metastasis remain unclear. We demonstrate the role of MT1-MMP on the lymph node metastasis using in vivo experimental model of lymph node metastasis by orthotopic implantation of MT1-MMP transfected gastric cancer cell line in the stomach of nude rats. TMK-1 cell line without expression of MT1-MMP was transfected with the pcDNA3 plasmid containing a 3.4-kb MT1-MMP cDNA fragment by calcium phosphate method, and the transfected cell line was designated as TMK-MT. Western blot and RT-PCR analyses showed the specific bands corresponding to MT1-MMP in the TMK-MT cells. By gelatin zymography, the activated form (62-kDa) of MMP-2 was identified in the medium of TMK-MT cell line, but was not detected in TMK-1 cells. Six weeks after orthotopic implantation of TMK-1 and TMK-MT xenografts of nude mouse-subcutaneus tumor into the stomach of nude rats, gastric tumors were found in all the animals. Histologically, the lymphatic invasion was found in the submucosa of the TMK-MT gastric tumors. Lymph node metastasis was not detected in nude rats bearing TMK-1 gastric tumor (0/8). In contrast, lymph node metastasis was detected in five out of 8 rats, bearing TMK-MT gastric tumor. MT1-MMP immunoreactivity was found on the cell membrane and cytoplasm of TMK-MT cells not only in the lymph node metastasis but also in the stomach tumor. These results suggest that MT1-MMP overexpression induced by transfection of its gene may promote lymph node metastasis of transformed cells.
Yonemura
Y
Y
Second Department of Surgery, School of Medicine, Kanazawa University, Japan. yonemu@med.kanazawa-u.ac.jp
Endo
Y
Y
Takino
T
T
Sakamoto
K
K
Bandou
E
E
Kinoshita
K
K
Fushida
S
S
Miwa
K
K
Sugiyama
K
K
Sasaki
T
T
D000818:Animals; D005260:Female; D008207:Lymphatic Metastasis; D053511:Matrix Metalloproteinase 14; D020778:Matrix Metalloproteinase 2 / Q000378:metabolism; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases / Q000502:physiology*; D051379:Mice; D008807:Mice, Inbred BALB C; D008819:Mice, Nude; D009363:Neoplasm Proteins / Q000502:physiology*; D009368:Neoplasm Transplantation; D051381:Rats; D011916:Rats, Inbred F344; D011923:Rats, Nude; D013047:Specific Pathogen-Free Organisms; D013274:Stomach Neoplasms / Q000473:pathology*; D014162:Transfection
C507096:Mmp14 protein, mouse; D009363:Neoplasm Proteins; D053504:Matrix Metalloproteinases, Membrane-Associated; D008666:Metalloendopeptidases; D020778:Matrix Metalloproteinase 2; D053511:Matrix Metalloproteinase 14