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Characterization of tissue factor expression on the human endothelial cell line ECV304
Author(s) -
LópezPedrera Chary,
Jardí Merce,
InglésEsteve Julia,
MuñozCánoves Pura,
Dorado Gabriel,
Velasco Francisco,
Félez Jordi
Publication year - 1997
Publication title -
american journal of hematology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 2.456
H-Index - 105
eISSN - 1096-8652
pISSN - 0361-8609
DOI - 10.1002/(sici)1096-8652(199710)56:2<71::aid-ajh1>3.0.co;2-y
Subject(s) - tissue factor , cell culture , endothelial stem cell , cell , microbiology and biotechnology , biology , human umbilical vein endothelial cell , umbilical vein , transcription factor , gene expression , phorbol , gene , signal transduction , medicine , biochemistry , in vitro , genetics , protein kinase c , coagulation
The endothelial cell line ECV304 is a spontaneously transformed cell line established from human umbilical vein. The characterization of tissue factor (TF) expression by ECV304 cells has been accomplished in this study. ECV304 cells expressed both TF mRNA and antigen (TF ag ) constitutively. In ECV304 cell lysates, the levels of TF ag (1.4 ± 0.3 ng of TF ag /10 6 cells) were considerably higher than in THP‐1 monocytoid cells (0.07 ± 0.03 ng of TF ag /10 6 cells). TF ag was also detected on the ECV304 cell surface by flow cytometric studies. In binding analyses, 3.5 ± 0.7 × 10 4 molecules of TF per cell were estimated, similar to the amounts found in ECV304 cell lysates (2.9 ± 0.6 × 10 4 molecules/cell), suggesting that all TF ag was translocated to the cell surface. Phorbol myristate acetate (PMA) stimulation of ECV304 cells resulted in an increase of TF mRNA levels, which was abrogated when gene transcription was impaired, suggesting a transcriptional regulation of the TF gene by PMA. In contrast, TF ag was not elevated by PMA‐stimulation, indicating the existence of additional posttranscriptional mechanisms. Thus, ECV304 cells constitute a singular endothelial cell model for exploring the regulation of TF expression. Am. J. Hematol. 56:71–78, 1997. © 1997 Wiley‐Liss, Inc.