PMID- 16919311
OWN - NLM
STAT- MEDLINE
DCOM- 20070614
LR  - 20070326
IS  - 0049-3848 (Print)
IS  - 0049-3848 (Linking)
VI  - 119
IP  - 6
DP  - 2007
TI  - Relative quantification of glycosaminoglycan-induced upregulation of TFPI-mRNA 
      expression in vitro.
PG  - 785-91
AB  - OBJECTIVE: Tissue factor pathway inhibitor (TFPI) is a multivalent Kunitz-type 
      serine proteinase inhibitor that plays a central role in the extrinsic pathway of 
      blood coagulation and is mainly expressed by endothelial cells. In this study we 
      examined the in vitro effects of heparin and other glycosaminoglycans on TFPI 
      mRNA-expression in cultivated human endothelial (Ea.hy 926) and in chondrosarcoma 
      (SW 1353) cells. METHODS: We used a LightCycler-based method for relative 
      quantification of the TFPI-mRNA expression before and after stimulation. The 
      cells were stimulated with different concentrations of heparin (with and without 
      addition of protamin), heparan sulfate (HS) and chondroitin-6-sulfate (CS). Cells 
      were harvested after incubation times of 4, 8 and 24h, total RNA was isolated, 
      and cDNA was synthesized and quantified relatively to a constantly expressed 
      housekeeping gene. RESULTS: Stimulation of Ea.hy 926 cells with unfractionated 
      heparin (UFH) or low-molecular-weight heparin (LMWH) caused a time- and 
      dose-dependent upregulation of TFPI-mRNA expression with LMWH showing the 
      stronger effect. In contrast to this, HS led to a strongly and CS to a slightly 
      decreased TFPI-mRNA expression. SW 1353 cells which were stimulated with LMWH/UFH 
      and HS/CS did not show a significant up- or downregulative effect. CONCLUSION: 
      Our results show that we have developed a versatile method for the relative 
      quantification of TFPI-mRNA expression. As a conclusion, the determined 
      heparin-induced upregulation of TFPI-mRNA expression can be considered a major 
      component of the modulation of the anticoagulant properties of the endothelium.
FAU - Thyzel, Elmar
AU  - Thyzel E
AD  - Institut fur Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum 
      Nordrhein-Westfalen, Universitatsklinik der Ruhr-Universitat Bochum, Bad 
      Oeynhausen, Germany.
FAU - Kohli, Sharmila
AU  - Kohli S
FAU - Siegling, Sabine
AU  - Siegling S
FAU - Prante, Christian
AU  - Prante C
FAU - Kleesiek, Knut
AU  - Kleesiek K
FAU - Gotting, Christian
AU  - Gotting C
LA  - eng
PT  - Journal Article
DEP - 20060817
PL  - United States
TA  - Thromb Res
JT  - Thrombosis research
JID - 0326377
RN  - 0 (Glycosaminoglycans)
RN  - 0 (Lipoproteins)
RN  - 0 (RNA, Messenger)
RN  - 0 (lipoprotein-associated coagulation inhibitor)
RN  - 9005-49-6 (Heparin)
RN  - 9007-28-7 (Chondroitin Sulfates)
RN  - 9050-30-0 (Heparitin Sulfate)
SB  - IM
MH  - Cell Line
MH  - Cell Proliferation/drug effects
MH  - Chondroitin Sulfates/pharmacology
MH  - Chondrosarcoma/*metabolism/pathology
MH  - Endothelial Cells/cytology/*metabolism
MH  - Glycosaminoglycans/*pharmacology
MH  - Heparin/pharmacology
MH  - Heparitin Sulfate/pharmacology
MH  - Humans
MH  - Lipoproteins/*genetics
MH  - RNA, Messenger/*metabolism
MH  - Up-Regulation/*drug effects
EDAT- 2006/08/22 09:00
MHDA- 2007/06/15 09:00
CRDT- 2006/08/22 09:00
PHST- 2005/09/17 00:00 [received]
PHST- 2006/04/09 00:00 [revised]
PHST- 2006/06/23 00:00 [accepted]
PHST- 2006/08/22 09:00 [pubmed]
PHST- 2007/06/15 09:00 [medline]
PHST- 2006/08/22 09:00 [entrez]
AID - S0049-3848(06)00232-5 [pii]
AID - 10.1016/j.thromres.2006.06.012 [doi]
PST - ppublish
SO  - Thromb Res. 2007;119(6):785-91. doi: 10.1016/j.thromres.2006.06.012. Epub 2006 
      Aug 17.