Characterization and functional analysis of TFPI-2 gene promoter in a human choriocarcinoma cell line.
Tissue factor pathway Inhibitor-2 (TFPI-2) is associated with extracellular matrices and plays a major role in cell migration and tumor invasion. In this study, a 4.8-kb human TFPI-2 gene 5'-flanking region was isolated, cloned and sequenced. Promoter region analysis revealed a high GC-rich content without canonical TATA and CAAT boxes but three transcription initiation sites were identified. Moreover, several putative binding sites for transcription factors were identified (MyoD, LYF1, NF-Y, GATA, oct-1, AP-1, Sp1, NF1, NF-kappa B and egr-1). To characterize potential regulatory regions, TFPI-2/luciferase promoter constructs were then transfected in human choriocarcinoma JEG-3 cells. We first showed that the minimal TFPI-2 promoter is located between -166 and -111 from the translation start site. Luciferase activity consistently increased after stimulation of JEG-3 cells by phorbol 12-myristate 13-acetate indicating that NF1, NF-kappa B and egr-1/Sp1 binding sites are crucial in inducible TFPI-2 expression. Moreover, negative regulatory regions included AP-1 binding sites were identified. This study demonstrates that the TFPI-2 gene promoter exhibits typical features of a housekeeping gene.
Hubé
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F
Laboratoire d'Hémostase, EA 3249 Cellules Hématopoíétiques, Hémostase et Greffe, IFR 120 Faculté de Médecine, 2 bis Bd Tonnellé, 37032 Tours Cedex, France.
Reverdiau
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P
Iochmann
S
S
Cherpi-Antar
C
C
Gruel
Y
Y
D024506:5' Flanking Region / Q000235:genetics; D001483:Base Sequence; D001665:Binding Sites; D045744:Cell Line, Tumor; D002822:Choriocarcinoma / Q000235:genetics / Q000473:pathology*; D003001:Cloning, Molecular; D005786:Gene Expression Regulation; D006023:Glycoproteins / Q000096:biosynthesis / Q000235:genetics*; D006801:Humans; D008969:Molecular Sequence Data; D011401:Promoter Regions, Genetic / Q000187:drug effects / Q000235:genetics*; D013755:Tetradecanoylphorbol Acetate / Q000031:analogs & derivatives* / Q000494:pharmacology; D014157:Transcription Factors; D024363:Transcription Initiation Site; D014162:Transfection
D006023:Glycoproteins; D014157:Transcription Factors; C021240:tissue-factor-pathway inhibitor 2; C016658:phorbolol myristate acetate; D013755:Tetradecanoylphorbol Acetate