Transcriptional silencing of the TFPI-2 gene by promoter hypermethylation in choriocarcinoma cells.
Tissue factor pathway inhibitor-2 (TFPI-2), a Kunitz-type serine proteinase inhibitor associated with the extracellular matrix, has been shown to reduce tumor invasion. In the present study we identified the presence of a complete CpG island region spanning exon 1 and the three transcription initiation sites. We demonstrate that DNA demethylation by 5'-aza-2'-deoxycytidine restores TFPI-2 transcription in JAR choriocarcinoma cells. The effect of in vitro DNA methylation on TFPI-2 promoter function was also confirmed with TFPI-2/luciferase promoter constructs. Finally, we determined the precise methylation status of CpG sites of the TFPI-2 promoter in normal and tumor trophoblast cells using the bisulfite genomic sequencing method. We conclude that hypermethylation of the TFPI-2 gene is correlated with transcriptional silencing and that the TFPI-2 gene may be a candidate tumor suppressor gene.
Hubé
Florent
F
Laboratoire d'Hémostase, EA 3249 'Cellules Hématopoïétiques, Hémostase et Greffe', INSERM U316, Faculté de Médecine, IFR 120 'Imagerie et Exploration fonctionnelles', 2bis Boulevard Tonnellé, F-37032 Tours cedex, France.
Reverdiau
Pascale
P
Iochmann
Sophie
S
Rollin
Jérôme
J
Cherpi-Antar
Catherine
C
Gruel
Yves
Y
D045744:Cell Line, Tumor; D002822:Choriocarcinoma; D018899:CpG Islands; D019175:DNA Methylation; D017931:DNA Primers; D004268:DNA-Binding Proteins / Q000378:metabolism; D004791:Enzyme Inhibitors / Q000494:pharmacology*; D020868:Gene Silencing; D006023:Glycoproteins / Q000235:genetics* / Q000378:metabolism; D006801:Humans; D008156:Luciferases / Q000235:genetics / Q000378:metabolism; D011401:Promoter Regions, Genetic; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014158:Transcription, Genetic
D017931:DNA Primers; D004268:DNA-Binding Proteins; D004791:Enzyme Inhibitors; D006023:Glycoproteins; C021240:tissue-factor-pathway inhibitor 2; D008156:Luciferases