Factor VIIa Induces Tissue Factor-dependent Up-regulation of Interleukin-8 in a Human Keratinocyte Line

Abstract

Tissue factor (TF), a transmembrane receptor for the serine protease coagulation factor VII(a) (FVIIa), is the main initiator of the coagulation cascade. Through incompletely elucidated mechanisms, TF serves additional functions in tumor-associated angiogenesis and metastasis. We have studied interleukin-8 (IL-8) as a possible link between TF-FVIIa complex formation and subsequent processes. Recombinant human FVIIa induced the up-regulation of both IL-8 mRNA and protein in a FVIIa dose-and time-dependent fashion. A neutralizing antibody to TF reduced this induction by 93 ؎ 5%. Active site-inhibited FVIIa had no stimulatory effect and completely blocked that of FVIIa. This confirms that the increased IL-8 production was dependent on the formation of TF-FVIIa complexes and the proteolytic activity of FVIIa. The IL-8 promoter contains DNA binding sites for nuclear factor-B (NF-B) and activator protein-1 (AP-1). In response to FVIIa, the DNA binding activity of both NF-B and AP-1 was enhanced in an electrophoretic mobility shift assay. In addition, the IL-8 promoter was transcriptionally activated both in a luciferase reporter system and a nuclear run-off assay. Moreover, IL-8 mRNA stability was significantly enhanced by FVIIa-induced activation of the mitogen-activated protein kinases ERK1/2 and p38. Taken together, TF-FVIIa signaling induced increased transcription as well as mRNA stabilization leading to the significant up-regulation of IL-8 protein synthesis.

Tid1 Negatively Regulates the Migratory Potential of Cancer Cells by Inhibiting the Production of Interleukin-8

“…4). Consistent with other studies (19,20), we found that the transcriptional activity of the mutated promoters was significantly reduced compared with the wild-type construct whether transfected with Tid1 siRNA oligos or not (Fig. 4).…”

“…The higher level of cellular Tid1 S (but not Tid1HQ) markedly blocked cell migration induced by FVIIa, which coincides with the ability of Tid1 S (but not Tid1HQ) to block FVIIa-induced IL-8 production in MDA-MB231 cells. Because NF-nB is the dominant pathway for the induction of IL-8 by FVIIa (20) and because Tid1 negatively regulates NF-nB activity (Fig. 4), it is possible that Tid1 blocked FVIIa-induced IL-8 production by inhibiting NF-nB activity.…”

“…The responsive DNA sequence for IL-8 induction is located in the IL-8 promoter between À133 and À85, and has been shown to contain the binding sites for transcriptional factors, AP-1, NF-nB, and C/EBP1 (18). It has been shown that NF-nB, AP-1, and to a lesser extent, C/EBP binding sites play a crucial role in controlling IL-8 promoter activity (19,20). These transcriptional factor binding sites in the IL-8 promoter have been individually or jointly mutated to keep them from binding their corresponding transcriptional factors, and the resultant mutated IL-8 promoters were placed separately in front of the luciferase reporter gene.…”

Tissue Factor-Dependent Chemokine Production Aggravates Experimental Colitis

“…Moreover, colon epithelial cells are a known source of IL-8 in inflammatory conditions (46), and the FVIIa/TF complex was previously shown to induce IL-8 levels in several different cell lines (47)(48)(49). In agreement, we show that ex vivo FVIIa-dependent KC production is dramatically reduced in TFlow colon sections compared with colon sections of wild-type mice.…”

Coagulation factors VIIa and Xa inhibit apoptosis and anoikis

“…Subsequent cellular events include the activation of p21Ras (Versteeg et al, 2003) and p42/p44 mitogen-activated protein kinase (MAP kinase) pathways (Poulsen et al, 1998;Camerer et al, 1999Camerer et al, , 2000bPendurthi et al, 2000;Versteeg et al, 2000). Furthermore, FVIIa induces the activation of the protein kinase B (PKB) pathway in TFexpressing cells, in turn producing enhanced protein synthesis (Versteeg et al, 2000(Versteeg et al, , 2002 and is also essential for FVIIa-induced production of IL-8 and other factors (Wang et al, 2002). Strikingly, both the p42/p44 MAP kinase and the PKB pathways are also known for their capacity to inhibit programmed cell death or apoptosis (Kennedy et al, 1997;Kulik et al, 1997;Bergmann et al, 1998;Kurada and White, 1998), but a possible link between TF and diminished apoptosis has not been investigated.…”