Abstract
L1 cell adhesion molecule (L1CAM) is a transmembrane cell adhesion molecule initially defined as a promigratory molecule in the developing nervous system that appears to be also expressed in some endothelial cells. However, little is known about the functional role of L1CAM on endothelial cells. We observed that L1CAM expression was selectively enhanced on endothelium associated with pancreatic adenocarcinoma in situ and on cultured pancreatic tumor-derived endothelial cells in vitro. L1CAM expression of endothelial cells could be augmented by incubation with immunomodulatory cytokines such as tumor necrosis factor alpha, interferon gamma, or transforming growth factor beta 1. Antibodies to L1CAM and the respective ligand neuropilin-1 blocked tube formation and stromal cell-derived factor 1β induced transmigration of tumor endothelial cells in vitro. L1CAM expression on tumor-derived-endothelial cells enhanced Panc1 carcinoma cell adhesion to endothelial cell monolayers and transendothelial migration. Our data demonstrate a functional role of L1CAM expression on tumor endothelium that could favor metastasis and angiogenesis during tumor progression.
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Abbreviations
- CFSE:
-
Carboxyfluoresceine diacete succinimidyl-ester
- EC:
-
endothelial cells
- HUVEC:
-
human umbilical vein endothelial cells
- HPMEC:
-
human pulmonary microvascular endothelial cells
- L1CAM:
-
L1 cell adhesion molecule
- NRP-1:
-
neuropilin-1
- Tu PAMEC:
-
tumor-derived pancreatic microvascular endothelial cells
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Acknowledgments
We thank Natalie Erbe and Maike Witt for excellent technical assistance. This work was supported by grants from Deutsche Krebshilfe to P.A. (Schwerpunktprogramm: Invasion and Migration) and the EU-FP6 framework program OVCAD project nr. PE-14034 to P.A.
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Supplemental Figure 1
Regulation of NRP-1 expression of cultured EC by proinflammatory cytokines. Flow cytometric analysis of NRP-1 expression on cultured non-malignant ECs (Con HUVEC, left, Con HPMEC, middle) and Tu PAMEC (right) untreated (dark gray histograms) or stimulated for 24 h with immunomodulatory cytokines TNF-α (400 U/ml, dotted line), IFN-γ (1,000 U/ml, dashed line), and TGF-β1 (10 ng/ml, dashed-dotted line). Light gray histograms represent negative control staining with respective secondary antibodies (GIF 12.4 KB).
Supplemental Figure 2
Analysis of conditions for achieving confluent endothelial monolayers. Hematoxylin and eosin staining of cultured non-malignant ECs (Con HUVEC) on gelatin-coated membranes. Different conditions (1 × 105/24 h, left panel and 2 × 105/48h, right panel) were tested to obtain a confluent EC monolayer. Original magnification ×400 (GIF 38.4 KB).
Supplemental Figure 3
L1CAM expression of Panc1 tumor cells. Flow cytometry analysis of L1CAM expression on cultured Panc1 cells (black line). Light gray histograms represent negative control staining with respective secondary antibody (GIF 4.83 KB).
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Issa, Y., Nummer, D., Seibel, T. et al. Enhanced L1CAM expression on pancreatic tumor endothelium mediates selective tumor cell transmigration. J Mol Med 87, 99–112 (2009). https://doi.org/10.1007/s00109-008-0410-7
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DOI: https://doi.org/10.1007/s00109-008-0410-7