TNF-alpha promotes lymphangiogenesis and lymphatic metastasis of gallbladder cancer through the ERK1/2/AP-1/VEGF-D pathway

Abstract

BackgroundTumor necrosis factor-alpha (TNF-α), a key player in cancer-related inflammation, was recently demonstrated to be involved in the lymphatic metastasis of gallbladder cancer (GBC). Vascular endothelial growth factor D (VEGF-D) is a key lymphangiogenic factor that is associated with lymphangiogenesis and lymph node metastasis in GBC. However, whether VEGF-D is involved in TNF-α-induced lymphatic metastasis of GBC remains undetermined.MethodsThe expression of VEGF-D in patient specimens was detected by immunohistochemistry and the relationship between VEGF-D in the tissue and TNF-α in the bile of the matching patients was analyzed. The VEGF-D mRNA and protein levels after treatment with exogenous TNF-α in NOZ, GBC-SD and SGC-996 cell lines were measured by real-time PCR and ELISA. The promoter activity and transcriptional regulation of VEGF-D were analyzed with the relative luciferase reporter assay, mutant constructs, electrophoretic mobility shift assay (EMSA), chromatin immunoprecipitation (ChIP) assay, RNA interference and Western blotting. Inhibitors of JNK, p38 MAPK and ERK1/2 were used to explore the upstream signaling effector of AP-1. We used lentiviral vector expressing a VEGF-D shRNA construct to knockdown VEGF-D gene in NOZ and GBC-SD cells. The role of the TNF-α-VEGF-D axis in the tube formation of human dermal lymphatic endothelial cells (HDLECs) was determined using a three-dimensional coculture system. The role of the TNF-α - VEGF-D axis in lymphangiogenesis and lymph node metastasis was studied via animal experiment.ResultsTNF-α levels in the bile of GBC patients were positively correlated with VEGF-D expression in the clinical specimens. TNF-α can upregulate the protein expression and promoter activity of VEGF-D through the ERK1/2 - AP-1 pathway. Moreover, TNF-α can promote tube formation of HDLECs, lymphangiogenesis and lymph node metastasis of GBC by upregulation of VEGF-D in vitro and in vivo.ConclusionTaken together, our data suggest that TNF-α can promote lymphangiogenesis and lymphatic metastasis of GBC through the ERK1/2/AP-1/VEGF-D pathway.

TNF-α promotes colon cancer cell migration and invasion by upregulating TROP-2

“…TNF-α may also upregulate MMPs in oral cancer cells, thereby promoting invasion and metastasis (18). In addition, TNF-α promotes lymphangiogenesis and lymphatic metastasis in gallbladder carcinoma by activating the ERK1/2 or the activator protein-1/vascular endothelial growth factor (VEGF)-D pathway (19). However, there are no previous reports describing the role of TNF-α in colon cancer invasion and metastasis.…”

PTTG1 is involved in TNF‐α‐related hepatocellular carcinoma via the induction of c‐myc

“…Accumulating evidence suggests that proinflammatory cytokines, including TNF‐α, IL‐6, IL‐1α, IL‐1β, and IFN‐γ, play a vital role in modulating autonomous growth signaling, which influences HCC growth and invasion, and TNF‐α is one of the most critical proinflammatory cytokines . TNF‐α plays a vital role in cancer by promoting the aberrant expression of a variety of genes through signaling pathway activation . The proinflammatory cytokine TNF‐α, which is mainly secreted by macrophages, can induce PTTG1 mRNA and protein expression in a dose‐dependent manner in keratinocytes .…”

“…7 TNF-α plays a vital role in cancer by promoting the aberrant expression of a variety of genes through signaling pathway activation. 35 The proinflammatory cytokine TNF-α, which is mainly secreted by macrophages, can induce PTTG1 mRNA and protein expression in a dose-dependent manner in keratinocytes. 9 Our study showed that TNF-α was upregulated in both HCC and paracancer tissues compared with normal liver tissues.…”

Tumor necrosis factor‐α induces prostate cancer cell migration in lymphatic metastasis through CCR7 upregulation

“…Increased C‐C chemokine receptor 7 (CCR7) expression in tumor cells has been reported to be significantly associated with lymph node metastasis in several cancer types . Interestingly, CCR7 has been found to mediate TNF‐α‐induced lymphatic metastasis of gallbladder cancer . C‐C chemokine receptor 7 is a receptor for the chemokine CCL21 and is expressed in various immune cells that migrate to and within lymphoid tissues.…”

“…[9][10][11] Interestingly, CCR7 has been found to mediate TNF-ainduced lymphatic metastasis of gallbladder cancer. 12,13 C-C chemokine receptor 7 is a receptor for the chemokine CCL21 and is expressed in various immune cells that migrate to and within lymphoid tissues. Chemokine CCL21 is abundantly expressed on fibroblastic reticular cells in lymph node metastasis.…”