Immunofluorescence and biochemical studies of the type VI collagen expression by human glioblastoma cells in vitro.
The human glioblastoma cell line U-87 MG was found to express a 140 kD polypeptide which was recognized on immunoblot analysis by a monoclonal antibody to type VI collagen. This polypeptide was digestible by a highly purified bacterial collagenase. After treatment of U-87 MG cells by pepsin, the protein profile revealed the two major pepsin-resistant fragments identical in Mr to those of collagen VI extracted from human placenta. The respective peptide maps from V8 protease one-dimensional gels of these two fragments were identical to those obtained with human collagen VI. Immunofluorescent staining by antibodies to type VI collagen was observed in the extracellular matrix. Moreover, U-87 MG cells were found to be positive for A2B5, a cell surface marker specific for O-2A type glial precursor cells. These data indicate that the human glioblastoma cell line U-87 MG exhibits the properties of glial precursor cells and expresses collagen type VI in vitro. This cell line therefore may prove valuable for comparative investigations of the regulation of type VI collagen synthesis, and may be useful as a model to study the function and pathological importance of type VI collagen in human brain tumours, both in vitro and in vivo.
Han
J
J
Department of Anatomy & Cell Biology, College of Medicine, University of Illinois, Chicago 60612.
Daniel
J C
JC
Lieska
N
N
Pappas
G D
GD
D015415:Biomarkers; D002460:Cell Line; D003094:Collagen / Q000032:analysis / Q000096:biosynthesis* / Q000302:isolation & purification; D004591:Electrophoresis, Polyacrylamide Gel; D005455:Fluorescent Antibody Technique; D005909:Glioblastoma / Q000378:metabolism*; D006801:Humans; D015151:Immunoblotting; D008970:Molecular Weight; D010434:Pepsin A; D010446:Peptide Fragments / Q000302:isolation & purification; D014407:Tumor Cells, Cultured
D015415:Biomarkers; D010446:Peptide Fragments; D003094:Collagen; D010434:Pepsin A