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Analysis of sequence variations in the promoter region of the human tissue factor pathway inhibitor 2 gene in apoplectic patients and blood donors

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Abstract

Tissue factor pathway inhibitor 2 (TFPI-2) is a Kunitz-type serine protease inhibitor with homology to TFPI-1, an important regulator of the extrinsic pathway of blood coagulation. Recent studies have focused on TFPI-2 and its implications for atherosclerosis. The promoter region and the exons of the human TFPI-2 gene were screened for sequence variations in 41 apoplectic patients and 140 blood donors with no history of ischemic stroke. The sequence variations −567T>C, −546T>C, −353A>G, −161G>C, −167G>A, −47C>A, and −18C>A, which are located in the TFPI-2 promoter, were discovered in both cohorts with allelic frequencies ranging from 0.3 to 2.4%. The influence of these sequence variations on the transcriptional activity of the TFPI-2 gene was investigated in HEK-293 cells using a promoter test system. A wild-type TFPI-2 promoter fragment 716 bp upstream of the translation start site was cloned into a secreted alkaline phosphatase expression vector, and the sequence variations were introduced by site-directed mutagenesis. Interestingly, the promoter activity of the tested mutants was reduced by 1.3- to 2.8-fold compared to that of wild-type control. The variation −18C>A, where a putative binding site of the transcription factor Sp-1 is located, had the strongest effect on transcriptional activity. In conclusion, our present study shows that the transcription of TFPI-2 is changed by single nucleotide polymorphisms and that the sequence variations in transcription factor binding sites of the TFPI-2 promoter may influence the regulation of this gene.

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Acknowledgement

We are grateful to Grainne Delany for linguistic advice and to the staff of the neurological department of the clinical center in Minden, Germany for blood drawing.

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Authors and Affiliations

  1. Institut für Laboratoriums- und Transfusionsmedizin, Herz- und Diabeteszentrum Nordrhein-Westfalen, Universitätsklinik der Ruhr-Universität Bochum, Georgstraße 11, 32545, Bad Oeynhausen, Germany

    Sabine Siegling, Elmar Thyzel, Knut Kleesiek & Christian Götting

  2. Klinik für Neurologie, Klinikum Minden, Minden, Germany

    Joerg Glahn

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  1. Sabine Siegling
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  2. Elmar Thyzel
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  4. Knut Kleesiek
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  5. Christian Götting
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Correspondence to Christian Götting.

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Siegling, S., Thyzel, E., Glahn, J. et al. Analysis of sequence variations in the promoter region of the human tissue factor pathway inhibitor 2 gene in apoplectic patients and blood donors. Ann Hematol 85, 32–37 (2006). https://doi.org/10.1007/s00277-005-0023-y

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  • DOI: https://doi.org/10.1007/s00277-005-0023-y

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