Nerve growth factor stimulates clonal growth of human lung cancer cell lines and a human glioblastoma cell line expressing high-affinity nerve growth factor binding sites involving tyrosine kinase signaling

• Published in: Cancer research (Chicago, Ill.) Vol. 55; no. 10; pp. 2212 - 2219
• Main Authors: Oelmann, E, Sreter, L, Schuller, I, Serve, H, Koenigsmann, M, Wiedenmann, B, Oberberg, D, Reufi, B, Thiel, E, Berdel, W E
• Format: Journal Article
• Language: English
• Published: United States 15.05.1995
• Subjects: Base Sequence; Brain Neoplasms - metabolism; Brain Neoplasms - pathology; Carcinoma, Non-Small-Cell Lung - metabolism; Carcinoma, Non-Small-Cell Lung - pathology; Carcinoma, Small Cell - metabolism; Carcinoma, Small Cell - pathology; Cell Differentiation - drug effects; Cell Division - drug effects; Dose-Response Relationship, Drug; Genistein; Glioblastoma - metabolism; Glioblastoma - pathology; Humans; Isoflavones - pharmacology; Lung Neoplasms - metabolism; Lung Neoplasms - pathology; Molecular Sequence Data; Nerve Growth Factors - pharmacology; Polymerase Chain Reaction; Protein-Tyrosine Kinases - antagonists & inhibitors; Protein-Tyrosine Kinases - physiology; Proto-Oncogene Proteins - metabolism; Receptor Protein-Tyrosine Kinases - metabolism; Receptor, trkA; Receptors, Nerve Growth Factor - metabolism; RNA-Directed DNA Polymerase; Tumor Cells, Cultured
• ISSN: 0008-5472

The growth of a panel of 22 different human tumor, leukemia, and lymphoma cell lines was examined in a human tumor cloning assay in agar or methylcellulose and a tritiated thymidine uptake assay. The cultures were performed in the absence or presence of increasing concentrations (0.5-500 ng/ml) of nerve growth factor (NGF). The growth of 17 of the 22 cell lines was not significantly and reproducibly affected by NGF. There was minor (1.2-fold) but reproducible stimulation of clonal growth in one glioblastoma cell line (86-HG-39) by NGF, but in this cell line NGF induced no growth modulation in a tritiated thymidine uptake assay. However, clonal growth of another glioblastoma cell line (87-HG-31) and all three lung cancer cell lines tested (HTB 119, HTB 120, CCL 185) could be stimulated up to 3-fold by NGF with a dose-response relationship for the growth factor. Growth stimulation by NGF could be completely reversed by neutralizing anti-NGF antibody and by the tyrosine kinase inhibitor genistein. Evaluation of secondary plating efficiency revealed the stimulation of colony formation as representing self-renewal and not terminal differentiation. Reverse transcriptase-PCR experiments in the five responding cell lines showed expression of both low-affinity NGF receptor (glycoprotein 75) and c-trk transcripts on the mRNA level. Of the five responding cell lines, only 86-HG-39, the cell line with the lowest responsiveness, revealed low-affinity NGF receptor on the protein level; the other four cell lines with high responsiveness, including the three lung cancer cell lines, expressed no low-affinity NGF receptor as shown by fluorescence-activated cell sorter analysis and immunoprecipitation using the ME 20.4 antibody. Immunoprecipitation using anti-trk antibodies was negative in all five responding cell lines. However, binding studies with iodinated NGF showed only low-affinity binding on the 86-HG-39 cell line and only high-affinity binding on the high-responder cell lines CCL 185 and 87-HG-31. In summary, our data suggest that NGF can be operative in stimulation of clonal growth of malignant tumor cells. High-affinity but not low-affinity binding sites mediate signal transduction for clonal growth and signaling involves tyrosine kinase activity.