Inhibition of TGF-beta-stimulated CTGF gene expression and anchorage-independent growth by cAMP identifies a CTGF-dependent restriction point in the cell cycle.
CTGF is a 38 kDa cysteine-rich peptide whose synthesis and secretion are selectively induced by transforming growth factor beta (TGF-beta) in connective tissue cells. We have investigated the signaling pathways controlling the TGF-beta induction of connective tissue growth factor (CTGF) gene expression. Our studies indicate that inhibitors of tyrosine kinases and protein kinase C do not block the signaling pathway used by TGF-beta to induce CTGF gene expression. In contrast, elevation of cAMP levels within the target cells by a variety of methods blocked the induction of CTGF by TGF-beta. Furthermore, agents that elevate cAMP blocked the induction of anchorage-independent growth (AIG) by TGF-beta. Inhibition of AIG could be overcome by the addition of CTGF, indicating that it was not a general inhibition of growth but a selective inhibition of CTGF synthesis that is responsible for the inhibition of TGF-beta-induced AIG by cAMP. Kinetic studies of the induction of DNA synthesis by CTGF in cells arrested by cAMP indicate that the block occurs in very late G1. These and other studies in monolayer cultures suggest that the CTGF restriction point in the cell cycle is distinct from the adhesion-dependent arrest point.
Kothapalli
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Department of Cell Biology and Anatomy, University of Miami School of Medicine, Florida 33136, USA.
Hayashi
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Grotendorst
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GR
D016475:3T3 Cells; D015124:8-Bromo Cyclic Adenosine Monophosphate / Q000494:pharmacology*; D000818:Animals; D016227:Benzoquinones; D002448:Cell Adhesion / Q000187:drug effects; D002453:Cell Cycle / Q000187:drug effects / Q000502:physiology*; D002455:Cell Division / Q000187:drug effects; D002460:Cell Line; D002772:Cholera Toxin / Q000494:pharmacology; D055513:Connective Tissue Growth Factor; D000242:Cyclic AMP / Q000378:metabolism*; D006152:Cyclic GMP / Q000378:metabolism; D005786:Gene Expression Regulation / Q000187:drug effects; D017930:Genes, Reporter; D006133:Growth Substances / Q000096:biosynthesis / Q000235:genetics* / Q000502:physiology; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D047029:Lactams, Macrocyclic; D008156:Luciferases / Q000096:biosynthesis / Q000235:genetics; D051379:Mice; D008934:Mitogens / Q000096:biosynthesis / Q000235:genetics; D016297:Mutagenesis, Site-Directed; D010982:Platelet-Derived Growth Factor / Q000494:pharmacology; D017354:Point Mutation; D016133:Polymerase Chain Reaction; D011809:Quinones / Q000494:pharmacology; D051381:Rats; D011993:Recombinant Fusion Proteins / Q000096:biosynthesis; D017828:Rifabutin / Q000031:analogs & derivatives; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014162:Transfection; D016212:Transforming Growth Factor beta / Q000494:pharmacology*
D016227:Benzoquinones; C526318:CCN2 protein, mouse; C526316:CCN2 protein, rat; D006133:Growth Substances; D017874:Immediate-Early Proteins; D036341:Intercellular Signaling Peptides and Proteins; D047029:Lactams, Macrocyclic; D008934:Mitogens; D010982:Platelet-Derived Growth Factor; D011809:Quinones; D011993:Recombinant Fusion Proteins; D016212:Transforming Growth Factor beta; D055513:Connective Tissue Growth Factor; D017828:Rifabutin; D015124:8-Bromo Cyclic Adenosine Monophosphate; C020754:herbimycin; D002772:Cholera Toxin; D000242:Cyclic AMP; D008156:Luciferases; D006152:Cyclic GMP; D013755:Tetradecanoylphorbol Acetate