E radiolabeled oligonucleotide PCP probe. Gem the result of EMSA has entered nicotine treatment Born a Erh Increase of AP1 dependent Independent transcriptional activity Transfected T cells in fa If the transition time with the AP reporter luciferase construction. Moreover, when transfected ECV304 cells that were transiently transfected with a decoy oligonucleotide AP 1, nicotine induces uPAR Promotoraktivit t decreased dose- Ngig by oligonucleotide-K Of. Was determined to determine whether the up-regulation of MAPK activation contributed to an AP nicotineinduced the effect of inhibitors of MAPK on AP-1 expression. Both PD98059 and SP600125 partially inhibited nicotine-induced AP-activity t. When the ECV304 cells PD98059 and SP600125 treated at a nicotineinduced AP activity t was also inhibited. The above results show that the transcription factor AP 1, which is activated by Erk 1/2 and JNK is induced in uPAR nicotine in human ECV304 cells are involved. The activation of NF B κ ROS by nicotine nicotine induced uPAR was examined to determine whether it is to activate the transcription factor NF B in human cells κ ECV304 k Nnten. To the R Of the transcription factor NF-B in κ uPAR expression induces nicotine, the effect of nicotine on the activation of NF B κ study was also investigated in ECV304 cells. EMSA and NF B-dependent Transcription κ Independent studies have shown that nicotine κ active NF B in a dose-dependent Ngigen way. The activation of NF B κ is usually associated with the induction of phosphorylation κ IB, which is followed by its degradation by the proteasome and NF B nuclear translocation κ. Thus, the Change in the amount of IB κ determined in ECV304 cells byWestern blotting using an antibody Were treated rpers against IB κ ECV304 cells with nicotine, showed a loss of the protein I B-κ a dose- Independent manner. The Hedgehog Signaling effects of BAY11 7082 on the mRNA expression by nicotine were then examined uPAR. As shown in Fig. 5C blocked by pretreatment nicotine 7082 BAY11-induced expression of uPAR mRNA. The involvement of NF B in the induction of uPAR κ by nicotine was best by co-transfection of ECV304 cells with the uPAR promoter-reporter and dominant-negative mutant forms of NF B κ related molecules CONFIRMS. As shown in Fig. 5D, expression of dominant negative mutant forms of IB I κ κ B or NIK has entered Born a decreased activity of t of nicotine-induced uPAR promoter. This suggests the involvement of NF B in κ uPAR induction by nicotine. Since nicotine is producing ROS and NF B, a redox-sensitive κ transcription factor, r Was responsible for the ROS in relation to NF-activation κ in the induction of uPAR nicotinemediated implicit. ECV304 cells were pretreated with NAC before treatment with nicotine and the effects of NAC on transcriptional activity of NF B t of κ were then examined. As shown in Fig. 5E, pretreatment of NAC blocked nicotine-induced NF-B activity t κ dependent Independent transcription in a dose-dependent Ngigen manner. These results show that intracellular Re H2O2 is produced by nicotine, contributed to the activation of NF B κ in human cells ECV304. Earlier reports have shown that nicotine induces ROS and activates the nucleotide Ren Etoposide transcription factor kappa B in cells ratmesencephalic. The involvement of ROS in endothelial cell invasion by nicotine is not defined yet. For this purpose.