Differential activation of the calcium/protein kinase C and the canonical beta-catenin pathway by Wnt1 and Wnt7a produces opposite effects on cell proliferation in PC12 cells.
We examined the effect of Wnt1 and Wnt7a on cell proliferation using undifferentiated PC12 cells, which originate from the neural crest and are widely employed as a neuronal cell model. Heterologous expression of Wnt1 enhanced [3H]thymidine incorporation and expression of cyclin D1 and cylin E in PC12 cells. Opposite effects were observed in PC12 cells expressing Wnt7a. Searching for the mechanisms underlying the opposite effects of Wnt1 and Wnt7a on PC12 cell proliferation, we examined the activation of the canonical beta-catenin/T-cell-lymphoid enhancer-binding protein transcription factor pathway and the 'calcium pathway' by co-transfecting the cells with a reporter gene controlled by either T-cell-lymphoid enhancer-binding protein transcription factor or the calcium-activated transcription factor, NFAT. Wnt1 and Wnt7a activated both pathways, but to a different extent. While Wnt1 preferentially activated the calcium pathway, Wnt7a mainly activated the canonical pathway. Pharmacological inhibition of protein kinase C, which is a component of the calcium pathway, abrogated the increase in cell proliferation induced by Wnt1 without affecting the antiproliferative action of Wnt7a. The action of Wnt7a was instead occluded by lithium ions, which mimic the activation of the canonical pathway, and was largely reduced by Dickkopf-1, which acts as an inhibitor of the canonical pathway. In addition, expression of a constitutively active mutant of beta-catenin potently activated the canonical Wnt pathway and reduced [3H]thymidine incorporation. These data challenge the view that the canonical Wnt pathway invariably supports cell growth and suggest that, at least in PC12 cells, cell proliferation is regulated by the balance between the calcium/protein kinase C pathway and the canonical pathway.
Spinsanti
Paola
P
Department of Human Physiology and Pharmacology, University of Rome La Sapienza, Rome, Italy.
De Vita
Teresa
T
Caruso
Alessandra
A
Melchiorri
Daniela
D
Misasi
Roberta
R
Caricasole
Andrea
A
Nicoletti
Ferdinando
F
D000818:Animals; D002118:Calcium / Q000378:metabolism; D002455:Cell Division / Q000502:physiology; D004742:Enhancer Elements, Genetic / Q000235:genetics; D004791:Enzyme Inhibitors / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D009474:Neurons / Q000166:cytology / Q000378:metabolism*; D016716:PC12 Cells; D011493:Protein Kinase C / Q000037:antagonists & inhibitors / Q000378:metabolism*; D011518:Proto-Oncogene Proteins / Q000235:genetics / Q000378:metabolism*; D051381:Rats; D015398:Signal Transduction / Q000502:physiology*; D013936:Thymidine / Q000493:pharmacokinetics; D014162:Transfection; D014316:Tritium; D051153:Wnt Proteins / Q000235:genetics / Q000378:metabolism*; D051155:Wnt1 Protein / Q000235:genetics / Q000378:metabolism*; D051176:beta Catenin / Q000378:metabolism*
D004791:Enzyme Inhibitors; D011518:Proto-Oncogene Proteins; D051153:Wnt Proteins; D051155:Wnt1 Protein; C495245:Wnt1 protein, rat; C490507:Wnt7a protein, rat; D051176:beta Catenin; D014316:Tritium; D008156:Luciferases; D011493:Protein Kinase C; D002118:Calcium; D013936:Thymidine