1. ADAM17 Promotes Motility, Invasion, and Sprouting of Lymphatic Endothelial Cells.
- Author
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Mężyk-Kopeć R, Wyroba B, Stalińska K, Próchnicki T, Wiatrowska K, Kilarski WW, Swartz MA, and Bereta J
- Subjects
- ADAM17 Protein, Cell Adhesion, Cell Line, Cell Proliferation, Cell Survival drug effects, Culture Media, ErbB Receptors metabolism, Extracellular Matrix Proteins metabolism, Gene Silencing, Humans, Integrins metabolism, Protein Kinase Inhibitors pharmacology, Receptors, Cell Surface metabolism, Signal Transduction, Transforming Growth Factor beta metabolism, Transforming Growth Factor beta2 metabolism, ADAM Proteins metabolism, Cell Movement, Endothelial Cells cytology, Endothelial Cells metabolism, Lymphangiogenesis
- Abstract
Tumor-associated lymphatic vessels actively participate in tumor progression and dissemination. ADAM17, a sheddase for numerous growth factors, cytokines, receptors, and cell adhesion molecules, is believed to promote tumor development, facilitating both tumor cell proliferation and migration, as well as tumor angiogenesis. In this work we addressed the issue of whether ADAM17 may also promote tumor lymphangiogenesis. First, we found that ADAM17 is important for the migratory potential of immortalized human dermal lymphatic endothelial cells (LEC). When ADAM17 was stably silenced in LEC, their proliferation was not affected, but: (i) single-cell motility, (ii) cell migration through a 3D Matrigel/collagen type I matrix, and (iii) their ability to form sprouts in a 3D matrix were significantly diminished. The differences in the cell motility between ADAM17-proficient and ADAM17-silenced cells were eliminated by inhibitors of EGFR and HER2, indicating that ADAM17-mediated shedding of growth factors accounts for LEC migratory potential. Interestingly, ADAM17 depletion affected the integrin surface expression/functionality in LEC. ADAM17-silenced cells adhered to plastic, type I collagen, and fibronectin faster than their ADAM17-proficient counterparts. The difference in adhesion to fibronectin was abolished by a cyclic RGD peptide, emphasizing the involvement of integrins in the process. Using a soluble receptor array, we identified BIG-H3 among several candidate proteins involved in the phenotypic and behavioral changes of LEC upon ADAM17 silencing. In additional assays, we confirmed the increased expression of BIG-H3, as well as TGFβ2 in ADAM17-silenced LEC. The antilymphangiogenic effects of ADAM17 silencing in lymphatic endothelial cells suggest further relevance of ADAM17 as a potential target in cancer therapy.
- Published
- 2015
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