title: Purification and identification of a novel and four known serine proteinase inhibitors secreted by human glioblastoma cells.
abstract: Our previous studies have shown that some human cancer cell lines produce pancreatic trypsinogen, plasminogen, and tissue-type kallikrein. To understand the regulatory mechanism of these proteinases, serine proteinase inhibitors secreted by human glioblastoma cell line T98G were analyzed by gelatin reverse zymography with trypsin. The serum-free conditioned medium of T98G cells showed more than ten trypsin inhibitor bands ranging from 16 to 150 kDa in the reverse zymography. Major trypsin inhibitors were purified by trypsin-affinity chromatography. Analysis of their N-terminal amino acid sequences demonstrated that the purified inhibitors were identical to the secreted forms of amyloid protein precursors (APPs), tissue factor pathway inhibitor (TFPI), placental protein 5 (PP5)/TFPI-2, and secretory leukocyte proteinase inhibitor (SLPI). In addition, a novel 25-kDa trypsin-binding protein, tentatively named p25TI, was identified. p25TI showed weak inhibitory activity against trypsin in reverse zymography as compared with the other inhibitors. The secretion of multiple forms of serine proteinase inhibitors by human cancer cells raises the possibility that they might be involved in the abnormal growth of cancer cells.
authors.affiliation: Division of Cell Biology, Yokohama City University.
authors.forename: N
authors.initials: N
authors.lastname: Koshikawa
authors.forename: T
authors.initials: T
authors.lastname: Nakamura
authors.forename: N
authors.initials: N
authors.lastname: Tsuchiya
authors.forename: M
authors.initials: M
authors.lastname: Isaji
authors.forename: H
authors.initials: H
authors.lastname: Yasumitsu
authors.forename: M
authors.initials: M
authors.lastname: Umeda
authors.forename: K
authors.initials: K
authors.lastname: Miyazaki
mesh_terms: D000595:Amino Acid Sequence; D001932:Brain Neoplasms / Q000378:metabolism*; D002850:Chromatography, Gel; D017077:Culture Media, Conditioned; D004591:Electrophoresis, Polyacrylamide Gel; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D008969:Molecular Sequence Data; D014361:Trypsin Inhibitors / Q000302:isolation & purification* / Q000378:metabolism; D014407:Tumor Cells, Cultured
chemical_list: D017077:Culture Media, Conditioned; D014361:Trypsin Inhibitors