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

Abstract

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 HDLECs 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 (ChIP). 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.

Transcription factors regulate GPR91-mediated expression of VEGF in hypoxia-induced retinopathy

“…Transcription factors control the expression of a given gene by serving as integration centers of different signaling cascades26. Many transcription factors regulate VEGF transcription and expression272829. Daft et al 27.…”

“…One report demonstrated that hyperglycemia significantly increased the binding of the transcription factor Sp1 to the VEGF-A promoter28. Du et al 29. used site-directed mutagenesis, electrophoretic mobility shift assay (EMSA) and ChIP technologies to determine the nuclear factor (NF)-κB binding site on the VEGF-C promoter.…”

The lymphatic system and pancreatic cancer

“…Such mediators include growth factors fibroblast growth factor-2 (FGF-2) [129], platelet-derived growth factor-BB (PDGF-BB) [130,131], nerve growth factor (NGF) [132], insulin-like growth factor-1 (IGF-1) and -2 [133], and hepatocyte growth factor (HGF) [134]; inflammatory cytokines interleukin-1 β (IL-1β) [135,136] and tumor necrosis factor- α (TNF-α) [135–137]; and other non-traditional signaling molecules lipid sphingosine-1-phosphate [138], cyclooxygenase 2 (Cox2), and EP3/4 [139]. A role for integrins in lymphangiogenesis is also emerging with evidence of binding of VEGF-A, -C, and -D to lymphatic endothelial-specific integrin-α9β1 [140,141].…”

“…While each of these factors does induce new lymphatic vessel growth, not all lymphangiogenesis is created equal; a study of corneal lymphangiogenesis in response to VEGF-A, VEGF-C, or FGF-2-loaded micropellets has revealed differences in both structure and function of lymphatic vessels and the proportion of blood to lymphatic vessels induced by these growth factors [142]. In the case of indirect stimulation of lymphangiogenesis, paracrine signals such as IL-1β [135,136] and TNF-α [135–137] can drive increased expression of the VEGFs, most notably VEGF-C. This likely occurs through activation of the NFκB promoter to induce VEGF-C expression [137].…”

“…In the case of indirect stimulation of lymphangiogenesis, paracrine signals such as IL-1β [135,136] and TNF-α [135–137] can drive increased expression of the VEGFs, most notably VEGF-C. This likely occurs through activation of the NFκB promoter to induce VEGF-C expression [137]. Other indirect inducers of lymphangiogenesis include Cox2 and EP3/4, which may increase expression of VEGF-C and -D to modulate cell growth during inflammation [139], and NGF, which increased expression of VEGF-C, but not VEGF-A, in a mouse corneal model of lymphangiogenesis [132].…”