Isolation and partial purification of growth factors with TGF-like activity from human malignant gliomas.
The effect of concentrated conditioned medium from each of eight human malignant glioma cell lines on the growth of indicator cells (normal rat kidney fibroblasts (NRK), clone 14) was determined in monolayer and in soft agar assay systems. The conditioned medium from all cell lines was mitogenic in the monolayer assay, but only SF-210, U-343 MG-A, and U-251 MG produced soluble factors that caused NRK cells to grow in soft agar. The soluble growth-promoting factors from these three cell lines were acid- and heat-stable (60 degrees C for 30 minutes) but were inactivated by trypsin (100 microns/ml) and dithiothreitol (50 microM). The growth factors from SF-210 and U-343 MG-A were further purified by molecular-sieve chromatography. The partially purified growth factor from U-343 MG-A retained transforming growth factor (TGF)-like activity, had a molecular weight of 9 kD, was potentiated by TGF-beta in the soft agar assay, competed effectively with 125I-epidermal growth factor (EGF) radiolabeled for the EGF receptor on A 431 epidermoid carcinoma cells, and was completely inhibited by monoclonal antibodies to TGF-alpha. The partially purified growth factor from SF-210 had a molecular weight of 17 kD, was not inhibited by monoclonal antibodies to platelet-derived growth factor (PDGF) or TGF-alpha, and did not bind to a heparin-Sepharose column. These results imply that U-343 MG-A secretes a growth factor with TGF-alpha-like activity, and SF-210 secretes a TGF with neither TGF-alpha nor TGF-beta activity.
Rutka
J T
JT
Department of Neurological Surgery, University of California, San Francisco.
Rosenblum
M L
ML
Stern
R
R
Ralston
H J
HJ
Dougherty
D
D
Giblin
J
J
DeArmond
S
S
D000362:Agar; D000818:Animals; D055598:Chemical Phenomena; D002627:Chemistry, Physical; D002850:Chromatography, Gel; D003470:Culture Media; D066246:ErbB Receptors / Q000032:analysis; D005347:Fibroblasts / Q000187:drug effects; D005910:Glioma / Q000032:analysis*; D006801:Humans; D007668:Kidney / Q000166:cytology; D015291:Transforming Growth Factors / Q000302:isolation & purification* / Q000494:pharmacology; D014407:Tumor Cells, Cultured
D003470:Culture Media; D015291:Transforming Growth Factors; D000362:Agar; D066246:ErbB Receptors