Physiological and chemical inducers of tissue factor pathway inhibitor-2 in human glioma cells.
Tissue factor pathway inhibitor-2 (TFPI-2), a serine protease inhibitor abundant in the extracellular matrix, is expressed in high amounts in low-grade, non-invasive glioma cells but in low amounts in high-grade, highly invasive glioma cells. Overexpression of TFPI-2 by highly invasive glioma cells reduces their invasiveness and thus may be useful in cancer therapy. The mechanisms underlying the transcriptional regulation of TFPI-2 are not well elucidated. We previously reported that the -312 to +1 region of TFPI-2 was critical for the minimal, inducible regulation of TFPI-2 in gliomas. This region harbors sites for several transcription factors, including SP1 (-192 to -183 and -135 to -128), AP-1 (-310 to -300, -213 to -204, and -163 to -154), NF-kappaB (-229 to -221), an NF-kappaB-like site (-291 to -281), and Lyf-1 (-260 to -252). Here we transiently transfected low-grade Hs683 glioma cells with mutant constructs to clarify the role of these transcription factors in TFPI-2 regulation. Addition of phorbol 12-myristate 13-acetate, 1,2-diacyl-sn-glycerol, IFN-gamma, or IFN-alpha induced the expression of TFPI-2 wild-type promoter construct as well as TFPI-2 protein and mRNA in Hs683 cells. Mutations at either of two AP-1 sites (-310 to -300 and -163 to -154) or either of two SP1 sites (-192 to -183 and -135 to -128) resulted in reduced TFPI-2 activity, regardless of the presence of stimulator compounds, and reduction in DNA-protein binding (by electrophoretic mobility shift assay).
Department of Biomedical and Therapeutic Sciences, Division of Cancer Biology, University of Illinois College of Medicine, Peoria, IL 61656, USA.
Santhi D
SD
Konduri
Niranjan
N
Yanamandra
Dzung H
DH
Dinh
William C
WC
Olivero
Meena
M
Gujrati
Donald C
DC
Foster
Walter
W
Kisiel
Jasti S
JS
Rao
D001483:Base Sequence; D002453:Cell Cycle / Q000187:drug effects; D017931:DNA Primers; D005786:Gene Expression Regulation / Q000187:drug effects / Q000235:genetics*; D005910:Glioma; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D016898:Interferon-alpha / Q000494:pharmacology; D007371:Interferon-gamma / Q000494:pharmacology; D008156:Luciferases / Q000235:genetics; D016133:Polymerase Chain Reaction; D011401:Promoter Regions, Genetic / Q000187:drug effects; D012333:RNA, Messenger / Q000235:genetics; D011994:Recombinant Proteins / Q000096:biosynthesis; D013755:Tetradecanoylphorbol Acetate / Q000494:pharmacology; D014157:Transcription Factors / Q000378:metabolism; D014158:Transcription, Genetic / Q000187:drug effects; D014162:Transfection; D014407:Tumor Cells, Cultured
D017931:DNA Primers; D006023:Glycoproteins; D016898:Interferon-alpha; D012333:RNA, Messenger; D011994:Recombinant Proteins; D014157:Transcription Factors; C021240:tissue-factor-pathway inhibitor 2; D007371:Interferon-gamma; D008156:Luciferases; D013755:Tetradecanoylphorbol Acetate