Journal of Southern Medical University ›› 2013, Vol. 33 ›› Issue (03): 351-.
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Abstract: Objective To explore the functional role of protein kinase D1 (PKD1) in the activation of nuclear factor-κB (NF-κB)signal pathway and NF-κB transcription mediated by Aspergillums fumigates. Methods A549 cells and HEK293 cells weretransfected with green fluorescence protein (GFP) or GFP-PKD1 followed by treatment with 1×105 CFU/ml Aspergillus fumigatusconidia for different time lengths. The phosphorylation levels of PKD1, IКB and p65 (pS276) in the transfected cells weremeasured by Western blotting. A549 cells were transfected with GFP-PKD1 or siRNA-PKD1, and the phosphorylation of IКBand p65 (pS276) was examined. Finally, NF-κB-luc and renilla luciferase reporter pRL-SV40 were cotransfected into GFP- orGFP-PKD1-transfected A549 cells before exposure of the cells to Aspergillus fumigatus conidia for 24 h, and NF-κBtranscriptional activity in the cells was determined using dual-luciferase reporter assay. Results Overexpression of PKD1significantly increased Aspergillus fumigatus conidia-stimulated phosphorylation of PKD1, IКB and p65 (pS276), whereas PKD1knockdown by siRNA-PKD1 suppressed IКB and p65 (pS276) phosphorylation. Dual luciferase assay demonstrated that PKD1overexpression markedly enhanced Aspergillus fumigatus-induced NF-κB transcription in A549 cells. Conclusion PKD1 maycontribute to the activation of NF-κB signal pathway and NF-κB transcription induced by Aspergillums fumigates.
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https://www.j-smu.com/EN/Y2013/V33/I03/351