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Abstract Background Tissue factor (TF) is a transmembrane protein that acts as a receptor for activated coagulation factor VII (FVIIa), initiating the coagulation cascade. Recent studies demonstrate that expression of tumor-derived TF also mediates intracellular signaling relevant to tumor growth and apoptosis. Our present study investigates the possible mechanism by which the interaction between TF and FVIIa regulates chemotherapy resistance in neuroblastoma cell lines. Methods Gene and siRNA transfection was used to enforce TF expression in a TF-negative neuroblastoma cell line and to silence endogenous TF expression in a TF-overexpressing neuroblastoma line, respectively. The expression of TF, Bcl-2, STAT5, and Akt as well as the phosphorylation of STAT5 and Akt in gene transfected cells or cells treated with JAK inhibitor and LY294002 were determined by Western blot assay. Tumor cell growth was determined by a clonogenic assay. Cytotoxic and apoptotic effect of doxorubicin on neuroblastoma cell lines was analyzed by WST assay and annexin-V staining (by flow cytometry) respectively. Results Enforced expression of TF in a TF-negative neuroblastoma cell line in the presence of FVIIa induced upregulation of Bcl-2, leading to resistance to doxorubicin. Conversely, inhibition of endogenous TF expression in a TF-overexpressing neuroblastoma cell line using siRNA resulted in down-regulation of Bcl-2 and sensitization to doxorubicin-induced apoptosis. Additionally, neuroblastoma cells expressing high levels of either endogenous or transfected TF treated with FVIIa readily phosphorylated STAT5 and Akt. Using selective pharmacologic inhibitors, we demonstrated that JAK inhibitor I, but not the PI3K inhibitor LY294002, blocked the TF/FVIIa-induced upregulation of Bcl-2. Conclusion This study shows that in neuroblastoma cell lines overexpressed TF ligated with FVIIa produced upregulation of Bcl-2 expression through the JAK/STAT5 signaling pathway, resulting in resistance to apoptosis. We surmise that this TF-FVIIa pathway may contribute, at least in part, to chemotherapy resistance in neuroblastoma.

This paper verifies the fluctuation on thermal parameters and ampacity of the high-voltage cross-linked polyethylene (XLPE) cables with different insulation conditions and describes the results of a thermal aging experiment on the XLPE insulation with different operating years in different laying modes guided by Comsol Multiphysics modeling software. The thermal parameters of the cables applied on the models are detected by thermal parameter detection control platform and differential scanning calorimetry (DSC) measurement to assure the effectivity of the simulation. Several diagnostic measurements including Fourier infrared spectroscopy (FTIR), DSC, X-ray diffraction (XRD), and breakdown field strength were conducted on the treated and untreated specimens in order to reveal the changes of properties and the relationship between the thermal effect and the cable ampacity. Moreover, a new estimation on cable ampacity from the perspective on XLPE insulation itself has been proposed in this paper, which is also a possible way to judge the insulation condition of the cable with specific aging degree in specific laying mode for a period of time. [ABSTRACT FROM AUTHOR]

Objective: Granulocyte macrophage colony-stimulating factor (GM-CSF) has been shown to specifically stimulate proliferation of CD34+ hematopoietic progenitor cells. Although signal transducers and activators of transcription 3 (STAT3) is believed essential for transduction of GM-CSF–induced cell proliferation, the signaling mediated by STAT3 is not completely understood. Because survivin regulates cell proliferation and survival via its antiapoptotic function, we studied the link between STAT3 signaling and survivin expression in CD34+ cells. Methods: GM-CSF–induced STAT3 and survivin expression in CD34+ cells was examined by Western blot assay. GM-CSF–activated survivin promoter activity was analyzed by gene transfection and reporter assays. The binding of STAT3 to the survivin promoter was evaluated by chromatin immunoprecipitation and electrophoretic mobility shift assay. Western blotting and flow cytometry were utilized to test the effect of Janus family of tyrosine kinases (JAK) inhibitor and STAT3 small interfering RNA (siRNA) on cell apoptosis. Results: We found that GM-CSF stimulates survivin promoter activity in CD34+ KG-1 cells, and STAT3 binds to the core survivin promoter containing a STAT response element TT(N)5AA at sites −264 to −256. Mutation or deletion of this STAT response element completely abolished the effects of GM-CSF on survivin promoter activity. Furthermore, addition of either JAK inhibitor or STAT3 siRNA was able to inhibit GM-CSF–induced survivin promoter activity and survivin expression. Inhibition of survivin by STAT3 siRNA or by withdrawal of GM-CSF in a GM-CSF–dependent, CD34+ line TF-1 decreased cell growth and increased apoptosis. Conclusion: Altogether, our results suggest that survivin is a transcriptional target of STAT3, and that GM-CSF–stimulated CD34+ cell proliferation is regulated by the JAK/STAT3/survivin signaling pathway. [Copyright &y& Elsevier]