Premium
T1α/Podoplanin through regulation of RhoA activation directs human lymphatic endothelial cell migration
Author(s) -
Navarro Angels,
Perez Ricardo E,
Rezaiekhaligh MO,
Mabry Sherry M,
Ekekezie Ikechukwu I
Publication year - 2009
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.23.1_supplement.1024.4
Subject(s) - rhoa , podoplanin , microbiology and biotechnology , lymphatic system , lymphatic endothelium , motility , wound healing , cell migration , lymphatic vessel , transfection , chemistry , cell , cancer research , biology , cell culture , signal transduction , medicine , immunology , biochemistry , genetics , cancer , metastasis
We have recently shown that early activation of RhoA in the lymphangiogenic process, which is required for the successful establishment of the capillary network, is dependent on podoplanin expression in human lung lymphatic microvascular endothelial cells HMVEC‐LLy ( Am J Physiol Lung Cell Mol Physiol 295: L543‐L551, 2008). We have also reported that podoplanin is required for a full migratory response to VEGF in these cells and that complete reorientation of the microtubule‐organizing center (MTOC) in response to VEGF in scratch‐wound assays, necessary for directional migration, is achieved only if the cells express podoplanin (FASEB J 2008; 22:1178.20). In the present study our objective was to determine if podoplanin regulates the migratory response of HMVEC‐LLy to VEGF via modulation of RhoA activation. RESULTS HMVEC‐LLy treated with CT04, a Rho inhibitor, achieve 60% wound closure by 48 hr in the presence of VEGF vs 100% in controls. Also, MTOC polarization in treated vs control cells is only 40% vs 80% as assesed at 3 hr after wounding. Furthermore, in transfection experiments, cells expressing a dominant negative mutant RhoA achieve only 20% wound closure vs 60% in control cells by 48 hr in the presence of VEGF. CONCLUSION Taken together our findings indicate that the migratory response of HMVEC‐LLy to VEGF stimulation is dependent on precise regulation of RhoA activity, a process dependent on podoplanin.