New human colorectal carcinoma cell lines that secrete proteinase inhibitors in vitro.

Abstract

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.

• Authors: H Kataoka, K Nabeshima, N Komada, M Koono
• Journal: Virchows Archiv. B, Cell pathology including molecular pathology (Virchows Arch B Cell Pathol Incl Mol Pathol) Vol. 57 Issue 3 Pg. 157-65 ( 1989) ISSN: 0340-6075 [Print] Germany
• PMID: 2570482 (Publication Type: Comparative Study, Journal Article, Research Support, Non-U.S. Gov't)

Chemical References

• Culture Media
• Protease Inhibitors
• Trypsin Inhibitors

Topics

• Adenocarcinoma (metabolism)
• Animals
• Chromatography, High Pressure Liquid
• Colorectal Neoplasms (metabolism)
• Culture Media
• Humans
• Immunodiffusion
• Immunohistochemistry
• Mice
• Mice, Nude
• Microscopy, Phase-Contrast
• Protease Inhibitors (metabolism)
• Trypsin Inhibitors (analysis, isolation & purification)
• Tumor Cells, Cultured (metabolism)