Negative growth regulation of SK-N-MC cells by bFGF defines a growth factor-sensitive point in G2.
Basic fibroblast growth factor (bFGF) has been shown to induce growth inhibition of the neuroepithelioma cell line SK-N-MC. Here we show that this growth inhibition occurs in G(2). We show that bFGF is active on these cells during S and early G(2) phase. Therefore, this constitutes a rather unusual mechanism of growth inhibition, because it is generally believed that cells become refractory to extracellular signals after passage through the restriction point. We show that bFGF treatment inhibits Tyr-15 dephosphorylation of cdc2 and prevents activation of Cdc25C, similar to what is seen upon activation of the G(2) DNA damage checkpoint. Interestingly, both DNA damage- and bFGF-induced effects on cdc2 phosphorylation are reverted by caffeine. To confirm the involvement of similar pathways induced by bFGF and DNA damage, we generated tetracycline-regulatable SK-N-MC clones expressing Cdc25C-S216A. Expression of this Cdc25C mutant can revert the bFGF-induced effects on cdc2 phosphorylation and can rescue cells from the block in G(2) imposed by bFGF. Taken together, these data define a growth factor-sensitive point in G(2) that most likely involves regulation of Cdc25C phosphorylation.
Smits
V A
VA
Jordan Laboratory, Department of Hematology, University Medical Center Utrecht G03.647, P.O. Box 85500, 3508 GA Utrecht, The Netherlands.
van Peer
M A
MA
Essers
M A
MA
Klompmaker
R
R
Rijksen
G
G
Medema
R H
RH
D002110:Caffeine / Q000494:pharmacology; D002455:Cell Division / Q000187:drug effects*; D019926:Cyclin B / Q000378:metabolism; D018844:Cyclin-Dependent Kinases / Q000378:metabolism; D016222:Fibroblast Growth Factor 2 / Q000494:pharmacology*; D016195:G2 Phase / Q000187:drug effects*; D006801:Humans; D010766:Phosphorylation; D014407:Tumor Cells, Cultured; D014443:Tyrosine / Q000378:metabolism
D019926:Cyclin B; D016222:Fibroblast Growth Factor 2; D002110:Caffeine; D014443:Tyrosine; D018844:Cyclin-Dependent Kinases