Reverse-zymographic analysis of protease nexin-II/amyloid beta protein precursor of human carcinoma cell lines, with special reference to the grade of differentiation and metastatic phenotype.
Trypsin inhibitors in serum-free conditioned media (SFCM) of various human carcinoma cell lines were analyzed by reverse zymography. Most of the cells secreted high-molecular-weight trypsin inhibitors (HMTI) larger than 100 kDa. The cell lines of colorectal carcinoma origin had a tendency to secrete HMTI whose molecular weight was a little higher than that of the other cell lines. Analysis of SFCM of subclones with different histological differentiation and metastatic/invasive potentials derived from a single pancreatic carcinoma cell line SUIT-2 showed that the HMTI activity in SFCM was correlated to the degree of histological differentiation in vivo and tended to be inversely correlated to their metastatic/invasive capabilities. Immunoblotting analysis revealed that these HMTI were protease nexin-II/amyloid beta protein precursors (PN-II/APP). Semi-quantificative reverse-transcriptase/polymerase-chain reaction study for PN-II/APP mRNAs suggested that the differences in PN-II/APP activities in SFCM between the subclones might be post-transcriptional or post-secretional events. In addition, SFCM of a highly metastatic subclone contained 43-kDa protein which reacted to anti-APP monoclonal antibody (MAb) suggesting that the subclone may have APP-degrading activity.
Kataoka
H
H
Second Department of Pathology, Miyazaki Medical College, Japan.
Seguchi
K
K
Iwamura
T
T
Moriyama
T
T
Nabeshima
K
K
Koono
M
M
D000230:Adenocarcinoma / Q000201:enzymology* / Q000473:pathology* / Q000556:secondary; D016564:Amyloid beta-Protein Precursor / Q000032:analysis*; D001483:Base Sequence; D002294:Carcinoma, Squamous Cell / Q000201:enzymology* / Q000473:pathology* / Q000556:secondary; D002295:Carcinoma, Transitional Cell / Q000201:enzymology* / Q000473:pathology* / Q000556:secondary; D002454:Cell Differentiation; D016895:Culture Media, Serum-Free; D006801:Humans; D008969:Molecular Sequence Data; D009362:Neoplasm Metastasis / Q000235:genetics; D009369:Neoplasms / Q000201:enzymology* / Q000473:pathology*; D010190:Pancreatic Neoplasms / Q000201:enzymology / Q000473:pathology; D010641:Phenotype; D015842:Serine Proteinase Inhibitors / Q000032:analysis; D014361:Trypsin Inhibitors / Q000032:analysis; D014407:Tumor Cells, Cultured
D016564:Amyloid beta-Protein Precursor; D016895:Culture Media, Serum-Free; D015842:Serine Proteinase Inhibitors; D014361:Trypsin Inhibitors