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Death receptor 6 (DR6) is required for mouse B16 tumor angiogenesis via the NF-κB, P38 MAPK and STAT3 pathways.

Yang X, Shi B, Li L, Xu Z, Ge Y, Shi J, Liu Y, Zheng D - Oncogenesis (2016)

Bottom Line: Although death receptor 6 (DR6) is aberrantly expressed in certain cancer cell lines, its function, signaling pathway and potential clinical significance in tumor progression are not well characterized.Deficiency of DR6 changes cytokine expression and secretion; in particular, it inhibits the proinflammatory cytokine interleukin-6 (IL-6), which is able to induce the expression of the angiogenesis-related factors: vascular endothelial growth factor-A, platelet-derived growth factor-β, vascular endothelial growth factor-D and platelet-derived growth factor receptor-α.Further experiments demonstrate that DR6-dependent angiogenesis is involved in the IL-6/P38 MAPK and IL-6/STAT3 pathways.

View Article: PubMed Central - PubMed

Affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

ABSTRACT
Although death receptor 6 (DR6) is aberrantly expressed in certain cancer cell lines, its function, signaling pathway and potential clinical significance in tumor progression are not well characterized. We report here that knocking down DR6 in the mouse B16 cell line has no effect on B16 cell death in vitro but suppresses xenograft B16 tumor growth by preventing tumor blood vessel formation in vivo. Deficiency of DR6 changes cytokine expression and secretion; in particular, it inhibits the proinflammatory cytokine interleukin-6 (IL-6), which is able to induce the expression of the angiogenesis-related factors: vascular endothelial growth factor-A, platelet-derived growth factor-β, vascular endothelial growth factor-D and platelet-derived growth factor receptor-α. Further experiments demonstrate that DR6-dependent angiogenesis is involved in the IL-6/P38 MAPK and IL-6/STAT3 pathways. Our novel findings demonstrate for the first time that DR6 expression in B16 cells facilitates tumor growth by accelerating tumor angiogenesis. Moreover, these results suggest that DR6 is involved in three important intracellular pathways that lead to homeostatic angiogenesis in tumor growth.

No MeSH data available.


Related in: MedlinePlus

DR6 mediates tumor angiogenesis through the NF-κB, IL-6/P38 MAPK and IL-6/STAT3 pathways. (a) Secreted IL-6 protein in B16 cell cultured media detected by ELISA. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with BAY or DMSO as a solvent control (C) for 12 h. (b) The levels of phosphorylated IκBα (p-IκBα) phosphorylated P38 (p-P38), phosphorylated STAT3 (p-STAT3), P38 and STAT3 were evaluated by western blot assays. (c) The detection of p-P38, total P38, p-STAT3 and total STAT3 proteins in DR6-deficient B16 cells by rescue experiment. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with IL-6 for 12 h. (d, e) The VEGF-D and PDGFR-α proteins in B16 cells lysate and VEGF-A protein in B16 cell cultured media detected by western blot or ELISA assays. (f) After 12 h treatment with DMSO, P38 inhibitor and STAT3 inhibitor, the secreted IL-6 protein levels from B16 cells with transfected siNC and siDR6 were examined by ELISA. (g) The secreted VEGF-A protein in the B16 cultivated media was examined by ELISA. The error bars represent the standard deviation of the mean values obtained from triplicate experiments. P<0.05 is considered statistically significant.
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fig5: DR6 mediates tumor angiogenesis through the NF-κB, IL-6/P38 MAPK and IL-6/STAT3 pathways. (a) Secreted IL-6 protein in B16 cell cultured media detected by ELISA. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with BAY or DMSO as a solvent control (C) for 12 h. (b) The levels of phosphorylated IκBα (p-IκBα) phosphorylated P38 (p-P38), phosphorylated STAT3 (p-STAT3), P38 and STAT3 were evaluated by western blot assays. (c) The detection of p-P38, total P38, p-STAT3 and total STAT3 proteins in DR6-deficient B16 cells by rescue experiment. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with IL-6 for 12 h. (d, e) The VEGF-D and PDGFR-α proteins in B16 cells lysate and VEGF-A protein in B16 cell cultured media detected by western blot or ELISA assays. (f) After 12 h treatment with DMSO, P38 inhibitor and STAT3 inhibitor, the secreted IL-6 protein levels from B16 cells with transfected siNC and siDR6 were examined by ELISA. (g) The secreted VEGF-A protein in the B16 cultivated media was examined by ELISA. The error bars represent the standard deviation of the mean values obtained from triplicate experiments. P<0.05 is considered statistically significant.

Mentions: IL-6 expression in the cultivated media from DR6-deficient B16 cells in the presence of NF-κB inhibitor BAY was largely reduced compared with control cells (Figure 5a), indicating that secreted IL-6 protein in DR6-deficient B16 cells is involved in NF-κB activation. B16 cells were treated with BAY can also prevent the expression IL-6, VEGF-A, PDGF-β mRNA and the VEGF-D and PDGFR-α protein (Supplementary Figure S7). Not surprisingly, IL-6 triggers P38 MAPK and STAT3 signaling in the wild-type B16 cells (Supplementary Figure S8).


Death receptor 6 (DR6) is required for mouse B16 tumor angiogenesis via the NF-κB, P38 MAPK and STAT3 pathways.

Yang X, Shi B, Li L, Xu Z, Ge Y, Shi J, Liu Y, Zheng D - Oncogenesis (2016)

DR6 mediates tumor angiogenesis through the NF-κB, IL-6/P38 MAPK and IL-6/STAT3 pathways. (a) Secreted IL-6 protein in B16 cell cultured media detected by ELISA. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with BAY or DMSO as a solvent control (C) for 12 h. (b) The levels of phosphorylated IκBα (p-IκBα) phosphorylated P38 (p-P38), phosphorylated STAT3 (p-STAT3), P38 and STAT3 were evaluated by western blot assays. (c) The detection of p-P38, total P38, p-STAT3 and total STAT3 proteins in DR6-deficient B16 cells by rescue experiment. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with IL-6 for 12 h. (d, e) The VEGF-D and PDGFR-α proteins in B16 cells lysate and VEGF-A protein in B16 cell cultured media detected by western blot or ELISA assays. (f) After 12 h treatment with DMSO, P38 inhibitor and STAT3 inhibitor, the secreted IL-6 protein levels from B16 cells with transfected siNC and siDR6 were examined by ELISA. (g) The secreted VEGF-A protein in the B16 cultivated media was examined by ELISA. The error bars represent the standard deviation of the mean values obtained from triplicate experiments. P<0.05 is considered statistically significant.
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Related In: Results  -  Collection

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fig5: DR6 mediates tumor angiogenesis through the NF-κB, IL-6/P38 MAPK and IL-6/STAT3 pathways. (a) Secreted IL-6 protein in B16 cell cultured media detected by ELISA. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with BAY or DMSO as a solvent control (C) for 12 h. (b) The levels of phosphorylated IκBα (p-IκBα) phosphorylated P38 (p-P38), phosphorylated STAT3 (p-STAT3), P38 and STAT3 were evaluated by western blot assays. (c) The detection of p-P38, total P38, p-STAT3 and total STAT3 proteins in DR6-deficient B16 cells by rescue experiment. B16 cells were transfected with siDR6 or control (siNC) for 48 h and then treated with IL-6 for 12 h. (d, e) The VEGF-D and PDGFR-α proteins in B16 cells lysate and VEGF-A protein in B16 cell cultured media detected by western blot or ELISA assays. (f) After 12 h treatment with DMSO, P38 inhibitor and STAT3 inhibitor, the secreted IL-6 protein levels from B16 cells with transfected siNC and siDR6 were examined by ELISA. (g) The secreted VEGF-A protein in the B16 cultivated media was examined by ELISA. The error bars represent the standard deviation of the mean values obtained from triplicate experiments. P<0.05 is considered statistically significant.
Mentions: IL-6 expression in the cultivated media from DR6-deficient B16 cells in the presence of NF-κB inhibitor BAY was largely reduced compared with control cells (Figure 5a), indicating that secreted IL-6 protein in DR6-deficient B16 cells is involved in NF-κB activation. B16 cells were treated with BAY can also prevent the expression IL-6, VEGF-A, PDGF-β mRNA and the VEGF-D and PDGFR-α protein (Supplementary Figure S7). Not surprisingly, IL-6 triggers P38 MAPK and STAT3 signaling in the wild-type B16 cells (Supplementary Figure S8).

Bottom Line: Although death receptor 6 (DR6) is aberrantly expressed in certain cancer cell lines, its function, signaling pathway and potential clinical significance in tumor progression are not well characterized.Deficiency of DR6 changes cytokine expression and secretion; in particular, it inhibits the proinflammatory cytokine interleukin-6 (IL-6), which is able to induce the expression of the angiogenesis-related factors: vascular endothelial growth factor-A, platelet-derived growth factor-β, vascular endothelial growth factor-D and platelet-derived growth factor receptor-α.Further experiments demonstrate that DR6-dependent angiogenesis is involved in the IL-6/P38 MAPK and IL-6/STAT3 pathways.

View Article: PubMed Central - PubMed

Affiliation: National Laboratory of Medical Molecular Biology, Institute of Basic Medical Sciences, Chinese Academy of Medical Sciences and Peking Union Medical College, Beijing, China.

ABSTRACT
Although death receptor 6 (DR6) is aberrantly expressed in certain cancer cell lines, its function, signaling pathway and potential clinical significance in tumor progression are not well characterized. We report here that knocking down DR6 in the mouse B16 cell line has no effect on B16 cell death in vitro but suppresses xenograft B16 tumor growth by preventing tumor blood vessel formation in vivo. Deficiency of DR6 changes cytokine expression and secretion; in particular, it inhibits the proinflammatory cytokine interleukin-6 (IL-6), which is able to induce the expression of the angiogenesis-related factors: vascular endothelial growth factor-A, platelet-derived growth factor-β, vascular endothelial growth factor-D and platelet-derived growth factor receptor-α. Further experiments demonstrate that DR6-dependent angiogenesis is involved in the IL-6/P38 MAPK and IL-6/STAT3 pathways. Our novel findings demonstrate for the first time that DR6 expression in B16 cells facilitates tumor growth by accelerating tumor angiogenesis. Moreover, these results suggest that DR6 is involved in three important intracellular pathways that lead to homeostatic angiogenesis in tumor growth.

No MeSH data available.


Related in: MedlinePlus