Characterization of platelet-derived growth factor (PDGF) action on a mouse neuroblastoma cell line, NB41, by introduction of an antisense PDGF beta-receptor RNA.
We have shown previously that platelet-derived growth factor (PDGF) has trophic effects on dopaminergic neurons in vitro. We now examined a mouse neuroblastoma cell line, NB41, for its response to PDGF and studied their phenotypic characteristics following introduction of an antisense PDGF beta-receptor RNA. NB41 cells produce both PDGF-AA and -BB; however, they carry only PDGF beta-receptors, responding to BB but not to AA. Culturing the cells with PDGF-BB induced mRNA for c-fos and PDGF-beta receptor as well as that of neuron-specific enzyme, tyrosine hydroxylase. In contrast, mRNA of chromogranin A, which is produced by chromaffin cells, decreased. Introduction of an antisense PDGF beta-receptor RNA in NB41 cells completely suppressed neurite extension and cell growth. We compared the PDGF-beta receptor sense and antisense clones for their survival. Following serum withdrawal, NB41 cells showed a DNA ladder, which by an addition of the neurotoxin, 6-hydroxy dopamine (6-OHDA), resulted in a further enhancement of the DNA ladder. The addition of PDGF-BB prior to 6-OHDA rescued cells from undergoing apoptosis, seen as a reduction of the DNA ladder. The antisense clone, regardless of the presence of PDGF-BB in the culture, showed a pronounced DNA ladder after serum withdrawal, which was further enhanced by the addition of 6-OHDA.
Funa
K
K
Ludwig Institute for Cancer Research, Biomedical Center, Uppsala, Sweden.
Ahgren
A
A
D000818:Animals; D017209:Apoptosis; D000077214:Becaplermin; D053379:Chromogranin A; D002864:Chromogranins / Q000235:genetics; D004273:DNA, Neoplasm / Q000032:analysis; D015972:Gene Expression Regulation, Neoplastic / Q000502:physiology*; D016762:Genes, fos / Q000235:genetics; D051379:Mice; D009447:Neuroblastoma / Q000235:genetics*; D009478:Neuropeptide Y / Q000235:genetics; D016627:Oxidopamine / Q000494:pharmacology; D010982:Platelet-Derived Growth Factor / Q000096:biosynthesis / Q000494:pharmacology*; D011494:Protein Kinases / Q000378:metabolism; D020574:Proto-Oncogene Proteins c-sis; D016372:RNA, Antisense / Q000494:pharmacology*; D012333:RNA, Messenger / Q000032:analysis; D020797:Receptor, Platelet-Derived Growth Factor beta; D017479:Receptors, Platelet-Derived Growth Factor / Q000032:analysis / Q000235:genetics / Q000502:physiology*; D014407:Tumor Cells, Cultured; D014446:Tyrosine 3-Monooxygenase / Q000235:genetics
D053379:Chromogranin A; D002864:Chromogranins; D004273:DNA, Neoplasm; D009478:Neuropeptide Y; D010982:Platelet-Derived Growth Factor; D020574:Proto-Oncogene Proteins c-sis; D016372:RNA, Antisense; D012333:RNA, Messenger; C075278:platelet-derived growth factor A; D000077214:Becaplermin; D016627:Oxidopamine; D014446:Tyrosine 3-Monooxygenase; D011494:Protein Kinases; D020797:Receptor, Platelet-Derived Growth Factor beta; D017479:Receptors, Platelet-Derived Growth Factor