H(2)O(2) induces upregulation of Fas and Fas ligand expression in NGF-differentiated PC12 cells: modulation by cAMP.
Fas, (APO-1/CD95), a transmembrane glycoprotein belonging to the tumor necrosis (TNF) receptor superfamily, transduces apoptotic death upon crosslinking by its cognate ligand (FasL). As upregulation of Fas/FasL expression occurs in neuropathological conditions (e.g., stroke, central nervous system [CNS] trauma and seizures) associated with oxidative damage, we questioned whether reactive oxygen species (ROS) can directly affect Fas and FasL expression in neuronal cells. Utilizing rat PC12 cells neuronally differentiated with nerve growth factor (NGF), we observed that concentrations of H(2)O(2) inducing apoptotic cell death rapidly trigger the expression of Fas mRNA and protein as well as FasL mRNA. Although NGF-addition to naive PC12 downregulated constitutive Fas and FasL transcription, the H(2)O(2)-induced Fas and FasL mRNA upregulation invariably occurred either in the presence or in the absence of NGF. Similarly, phorbol 1,2-myristate 1, 3-acetate (PMA), a potent protein kinase C (PKC) activator, did not modify Fas and FasL mRNA upregulation subsequent to H(2)O(2) exposure. On the contrary, forskolin and dibutyryl cAMP, which elevate intracellular cAMP by independent mechanisms, both counteracted H(2)O(2)-induced Fas, but not FasL, mRNA upregulation and increased constitutive expression of FasL mRNA. Altogether, our data show that oxidative stress is a major stimulus in eliciting Fas and FasL expression in NGF-differentiated PC12 cells. Moreover, we describe here for the first time the existence of cAMP-dependent mechanism(s) modulating Fas and FasL expression.
Research and Innovation Company, Padova, Italy. facchinetti@researchinnovation.com
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Facchinetti
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Furegato
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D000818:Animals; D016923:Cell Death; D000242:Cyclic AMP / Q000378:metabolism*; D053938:DNA Fragmentation; D053222:Fas Ligand Protein; D006861:Hydrogen Peroxide / Q000009:adverse effects*; D008562:Membrane Glycoproteins / Q000187:drug effects / Q000378:metabolism*; D020932:Nerve Growth Factor / Q000378:metabolism*; D016877:Oxidants / Q000009:adverse effects*; D018384:Oxidative Stress; D016716:PC12 Cells; D012333:RNA, Messenger / Q000032:analysis; D051381:Rats; D020133:Reverse Transcriptase Polymerase Chain Reaction; D015854:Up-Regulation / Q000187:drug effects; D019014:fas Receptor / Q000187:drug effects / Q000378:metabolism*
D053222:Fas Ligand Protein; C505662:Faslg protein, rat; D008562:Membrane Glycoproteins; D016877:Oxidants; D012333:RNA, Messenger; D019014:fas Receptor; D020932:Nerve Growth Factor; D006861:Hydrogen Peroxide; D000242:Cyclic AMP