Transendothelial Migration of Colon Carcinoma Cells Requires Expression of E-selectin by Endothelial Cells and Activation of Stress-activated Protein Kinase-2 (SAPK2/p38) in the Tumor Cells

Abstract

Adhesion and migration of tumor cells on and through the vascular endothelium are critical steps of the metastatic invasion. We investigated the roles of E-selectin and of stress-activated protein kinase-2 (SAPK2/p38) in modulating endothelial adhesion and transendothelial migration of HT-29 colon carcinoma cells. Tumor necrosis factor ␣ (TNF␣) strongly increased the expression of E-selectin in human umbilical vein endothelial cells (HUVEC). This effect was independent of the activation of SAPK2/p38 induced by TNF␣. Adhesion of HT-29 cells on a monolayer of HUVEC pretreated with TNF␣ was dependent on E-selectin expression but was independent of SAPK2/p38 activity of both HUVEC and tumor cells. The adhesion of HT-29 cells to E-selectin-expressing HUVEC led to the activation of SAPK2/p38 in the tumor cells as reflected by the increased phosphorylation of the actin-polymerizing factor HSP27 by mitogenactivated protein kinase 2/3, a direct target of SAPK2/ p38. Moreover, a recombinant E-selectin/Fc chimera quickly increased the activation of SAPK2/p38 in HT-29 cells. Blocking the increased activity of SAPK2/p38 of HT-29 cells by SB203580 or by expressing a dominant negative form of SAPK2/p38 inhibited their transendothelial migration. Similarly, HeLa cells stably expressing a kinase-inactive mutant of SAPK2/p38 showed a decreased capacity to cross a layer of HUVEC. Overall, our results suggest that the regulation of transendothelial migration of tumor cells involves two essential steps as follows: adhesion to the endothelium through adhesion molecules, such as E-selectin, and increased motogenic potential through adhesion-mediated activation of the SAPK2/p38 pathway.Circulating tumor cells attach to adhesive endothelial molecules, and these interactions are pivotal during the metastatic process. E-selectin, whose expression is induced by cytokines and growth factors released by tumor cells, promotes the endothelial adhesion of tumor cells from various origins, and this correlates with metastatic dissemination of tumor cells, e.g. to liver, lung, and bones (1-4). The ability of colon tumor cell clones to bind E-selectin on endothelial cells is even directly proportional to their metastatic potential (5). Moreover, inhibiting the expression of E-selectin with drugs such as cimetidine prevents metastasis (6). Metastatic colonization also correlates with the expression of other types of endothelial adhesion molecules such as P-selectin and ICAM 1 (7-12). Furthermore, the metastatic potential is associated with the circulating levels of soluble endothelial adhesion molecules shed by activated endothelial cells of cancer patients (13-17). The increased metastatic potential associated with adhesion of tumor cells to the endothelium might result from two distinctive processes as follows: local intravascular proliferation of the attached tumor cells or extravasation of these cells following their transendothelial migration into the sub-vascular tissues (18,19). In both cases, the underlying biochemical mechanisms remain ill-...

Regulation of transendothelial migration of colon cancer cells by E-selectin-mediated activation of p38 and ERK MAP kinases

“…Activation of E-selectin by crosslinking antibodies or adhesion of colon cancer cells triggers the activation of ERK and p38 MAP kinases in endothelial cells Transendothelial migration of colon cancer cells depends on motogenic signals that result from their adhesion to E-selectin expressed by endothelial cells (Laferrie`re et al, 2001). In the present work, we investigated whether the adhesion of colon cancer cells to E-selectin also induced in endothelial cells a forward signalling that leads to the formation of breaches in the endothelial layer, thereby facilitating invasiveness.…”

“…It requires specific interactions between adhesion receptors present on vascular endothelial cells and their counter-receptors on cancer cells (Haier and Nicolson, 2001). These adhesive interactions induce a reverse signalling in the cancer cells that increases their motile potential and a forward signalling in endothelial cells that elicits breaches in the endothelium layer (Hu et al, 2000;Laferrie`re et al, 2001;Weis et al, 2004). However, in both cases the signalling mechanisms involved are still ill defined.…”

Regulation of Sialyl Lewis Antigen Expression in Colon Cancer Cells by Sialidase NEU4

“…To clarify details with reference to signal transduction, we focused attention on the p38/Hsp27 pathway. It is known that binding of sialyl-Le a and sialyl-Le x to E-selectin activates p38 signaling in cancer cells (12,39), resulting in stimulation of phosphorylation of various transcription factors and MAPKAP kinase 2, with subsequent activation of MAPKAP kinase 2 and Hsp27, the actin polymerizing factor (40). To investigate the influence of NEU4S on cell motility, HT29 cells were stimulated with E-selectin and the phosphorylation levels were assessed.…”

“…In normal colon mucosa, the expression of sialyl-Le a and sialyl-Le x is quite low, whereas more complicated carbohydrates, such as disialyl-Le a and sialyl-6-sulfo-Le x , are major components (9,10). Sialyl-Le a and sialyl-Le x play an important role, as carbohydrate ligands for E-selectin, in E-selectin-mediated cancer cell adhesion to vascular endothelial cells during the course of hematogenous metastasis (11), through activation of the SAPK/p38 signaling pathway (12,13). On the other hand, disialyl-Le a , sialyl-6-sulfoLe x , Le a , and Le x do not bind to E-selectin.…”

Cimetidine in colorectal cancer – are the effects immunological or adhesion-mediated?

“…SL X expression is increased in human hepatic colorectal metastases compared to the primary tumour (Hoff et al, 1989) and expression has been shown to correlate with poor survival of colorectal cancer patients (for example, in stage III patients 5 year survival was 42% in the sL X positive group vs 81% in patients with tumours negative for sL X in one study) (Nakamori et al, 1993). Expression of E-selectin on endothelial cells has also recently been shown to be essential in models of transendothelial migration of colon carcinoma cells (Laferriere et al, 2001). Matsumoto et al (2001) have also retrospectively analyzed the patients in their previous study of cimetidine in colorectal cancer according to sL A and sL X expression.…”