Inhibition of PLC-γ1 activity converts nerve growth factor from an anti-mitogenic to a mitogenic signal in CHO cells
Nerve growth factor (NGF) treatment of Chinese hamster ovary fibroblast (CHO) cells exogenously expressing 2.5–105 TrkA receptors (CHO/TrkA) results in inhibition of serum and insulin-like growth factor-I (IGF-I) stimulated cell proliferation in a dose-dependent manner. Furthermore, NGF does not sti...
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Published in | Oncogene Vol. 18; no. 35; pp. 4908 - 4919 |
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Main Authors | , , , |
Format | Journal Article |
Language | English |
Published |
Basingstoke
Nature Publishing
02.09.1999
Nature Publishing Group |
Subjects | |
Online Access | Get full text |
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Summary: | Nerve growth factor (NGF) treatment of Chinese hamster ovary fibroblast (CHO) cells exogenously expressing 2.5–105 TrkA receptors (CHO/TrkA) results in inhibition of serum and insulin-like growth factor-I (IGF-I) stimulated cell proliferation in a dose-dependent manner. Furthermore, NGF does not stimulate [3H]thymidine incorporation and inhibits IGF-I mediated DNA synthesis in CHO/TrkA cells. NGF and IGF-I induce extracellular-signal regulated kinase 1 (ERK1) and ERK2 activation, but NGF is able to stimulate a higher and more sustained activation of these enzymes compared with IGF-I. Cotreatment with NGF and IGF-I yields an ERK1/2 activity profile similar to that of NGF treatment alone. While pretreatment with mitogen activated protein kinase kinase (MKK) inhibitor PD98059 (30 μM) results in 100% inhibition of IGF-I stimulated MAPK phosphorylation (IC50<1 μM), NGF mediated MAPK phosphorylation is only decreased by 50% (IC50=3 μM). NGF, but not IGF-I, stimulates tyrosine phosphorylation and activation of PLC-γ1 which can be inhibited in a dose-dependent manner by phosphoinositide-specific phospholipase C (PI-PLC) inhibitor U73122 (IC50=4 μM). Pretreatment with U73122 (IC50=7 μM) results in an 87% inhibition of NGF mediated MAPK phosphorylation, while cotreatment with PD98059 and U73122 results in 97% inhibition. U73122 pretreatment has no effect on NGF stimulated Akt activation. NGF, but not IGF-I, stimulates the tyrosine phosphorylation of Suc1-associated neurotrophic factor-induced tyrosine phosphorylation target (SNT-1)/fibroblast growth factor receptor substrate 2 (FRS2) which can be completely prevented by pretreatment with 10 μM U73122. Finally, inhibition of PI-PLC results in NGF's ability to stimulate DNA synthesis in the absence and presence of IGF-I. |
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ISSN: | 0950-9232 1476-5594 |
DOI: | 10.1038/sj.onc.1202861 |