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Vascular endothelial growth factor ( VEGF )‐C is an important lymphangiogenic factor involved in the lymphangiogenesis of gallbladder carcinoma ( GBC ) and the lymph node metastasis of the tumor. Tumor necrosis factor ( TNF )‐α, a key inflammatory cytokine responding to chronic inflammation of  GBC , has been reported to stimulate the expression of  VEGF ‐C in some nonneoplastic cells. But whether  TNF ‐α promotes the expression of  VEGF ‐C in  GBC  has yet to be determined. Therefore, in the present study, the concentration of  TNF ‐α and  VEGF ‐C and the lymphatic vessel density ( LVD ) in the clinical  GBC  specimens were analyzed, and a linear correlation was found between the concentration of  TNF ‐α and that of  VEGF ‐C, the lymphatic vessel density ( LVD ); The transcription and protein level of  VEGF ‐C in  NOZ  cell line were detected by real‐time polymerase chain reaction ( PCR ) and enzyme linked immunosorbent assay ( ELISA ), and  TNF ‐α enhanced the expression of  VEGF ‐C in  NOZ  cell lines in a dose and time‐dependent manner. Lymphatic tube formation  in vitro  was observed in a three‐dimensional coculture system consisting of  HDLEC s and  NOZ  cell lines, and lymphatic vessels of  GBC  in nude mice model was detected by immunohistochemistry.  TNF ‐α promoted the tube formation of lymphatic endothelial cells  in vitro  and the lymphangiogenesis of  GBC  in nude mice; The nuclear factor ( NF )‐κB binding site on the  VEGF ‐C promoter was identified using Site‐directed mutagenesis, electrophoretic mobility shift assay ( EMSA ) and chromatin immunoprecipitation assay (Ch IP ). Taken together,  TNF ‐α can upregulate the expression of  VEGF ‐C and promote the lymphangiogenesis of  GBC  via  NF ‐κB combining with the promoter of  VEGF ‐C.

cancer science

Tumor necrosis factor‐α promotes the lymphangiogenesis of gallbladder carcinoma through nuclear factor‐κB‐mediated upregulation of vascular endothelial growth factor‐C