New human colorectal carcinoma cell lines that secrete proteinase inhibitors in vitro.
Two new human cell lines, RCM-1 and CoCM-1, have been established from primary colorectal adenocarcinomas. Both cell lines were unique in that the cultures secreted trypsin inhibitors in vitro. The activities of these inhibitors were accumulated in serum-free media of both cell lines over a period of several days. Two inhibitors (PI-1 and PI-2) were isolated from serum-free conditioned medium in which RCM-1 was grown by anion-exchange and gel filtration high-performance liquid chromatography. PI-1 inhibited trypsin and chymotrypsin strongly, and pancreatic elastase weakly. Its molecular weight was about 57 kilodaltons (Kd) as determined by gel filtration chromatography. It cross-reacted with the antiserum elicited against human alpha 1-antitrypsin in double immunodiffusion. PI-1 corresponding to alpha 1-antitrypsin was also demonstrated immunohistochemically in both cell lines. PI-2 inhibited trypsin strongly, and chymotrypsin, kallikrein and plasmin weakly. It had higher molecular weight (200-300 Kd) than that of PI-1, and did not cross-react with antisera against human alpha 1-antitrypsin, alpha 2-macroglobulin, alpha 1-antichymotrypsin, alpha 2-plasmin inhibitor, inter-alpha-trypsin inhibitor and urinary trypsin inhibitor. RCM-1 and CoCM-1 are the first colorectal adenocarcinoma cell lines that secrete functionally active trypsin inhibitors, including alpha 1-antitrypsin in vitro, and are useful for the study of tumor-cell derived proteinase inhibitors.
Kataoka
H
H
Department of Pathology, Miyazaki Medical College, Japan.
Nabeshima
K
K
Komada
N
N
Koono
M
M
D000230:Adenocarcinoma / Q000378:metabolism*; D000818:Animals; D002851:Chromatography, High Pressure Liquid; D015179:Colorectal Neoplasms / Q000378:metabolism*; D003470:Culture Media; D006801:Humans; D005779:Immunodiffusion; D007150:Immunohistochemistry; D051379:Mice; D008819:Mice, Nude; D008858:Microscopy, Phase-Contrast; D011480:Protease Inhibitors / Q000378:metabolism*; D014361:Trypsin Inhibitors / Q000032:analysis / Q000302:isolation & purification; D014407:Tumor Cells, Cultured / Q000378:metabolism
D003470:Culture Media; D011480:Protease Inhibitors; D014361:Trypsin Inhibitors