Tissue factor pathway inhibitor-2 was repressed by CpG hypermethylation through inhibition of KLF6 binding in highly invasive breast cancer cells

Hongshen Guo, Yifeng Lin, Hongwei Zhang, Juan Liu, Nong Zhang, Yiming Li, Desheng Kong, Qiqun Tang, Duan Ma

Research output: Contribution to journalArticle

Abstract

Background: Tissue factor pathway inhibitor-2 (TFPI-2) is a matrix-associated Kunitz inhibitor that inhibits plasmin and trypsin-mediated activation of zymogen matrix metalloproteinases involved in tumor progression, invasion and metastasis. Here, we have investigated the mechanism of DNA methylation on the repression of TFPI-2 in breast cancer cell lines. Results: We found that both protein and mRNA of TFPI-2 could not be detected in highly invasive breast cancer cell line MDA-MB-435. To further investigate the mechanism of TFPI-2 repression in breast cancer cells, 1.5 Kb TFPI-2 promoter was cloned, and several genetic variations were detected, but the promoter luciferase activities were not affected by the point mutation in the promoter region and the phenomena was further supported by deleted mutation. Scan mutation and informatics analysis identified a potential KLF6 binding site in TFPI-2 promoter. It was revealed, by bisulfite modified sequence, that the CpG island in TFPI-2 promoter region was hypermethylated in MDA-MB-435. Finally, using EMSA and ChIP assay, we demonstrated that the CpG methylation in the binding site of KLF-6 diminished the binding of KLF6 to TFPI-2 promoter. Conclusion: In this study, we found that the CpG islands in TFPI-2 promoter was hypermethylated in highly invasive breast cancer cell line, and DNA methylation in the entire promoter region caused TFPI-2 repression by inducing inactive chromatin structure and decreasing KLF6 binding to its DNA binding sequence.

Original languageEnglish (US)
Article number110
JournalBMC Molecular Biology
Volume8
DOIs
StatePublished - Dec 3 2007
Externally publishedYes

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Cells
Breast Neoplasms
Genetic Promoter Regions
CpG Islands
DNA Methylation
Cell Line
Binding Sites
tissue-factor-pathway inhibitor 2
Antifibrinolytic Agents
Mutation
Enzyme Precursors
Informatics
Methylation
Trypsin Inhibitors
Fibrinolysin
Luciferases
Matrix Metalloproteinases
Point Mutation
Trypsin
Chromatin

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)
  • Biochemistry

Cite this

Tissue factor pathway inhibitor-2 was repressed by CpG hypermethylation through inhibition of KLF6 binding in highly invasive breast cancer cells. / Guo, Hongshen; Lin, Yifeng; Zhang, Hongwei; Liu, Juan; Zhang, Nong; Li, Yiming; Kong, Desheng; Tang, Qiqun; Ma, Duan.

In: BMC Molecular Biology, Vol. 8, 110, 03.12.2007.

Research output: Contribution to journalArticle

Guo, Hongshen ; Lin, Yifeng ; Zhang, Hongwei ; Liu, Juan ; Zhang, Nong ; Li, Yiming ; Kong, Desheng ; Tang, Qiqun ; Ma, Duan. / Tissue factor pathway inhibitor-2 was repressed by CpG hypermethylation through inhibition of KLF6 binding in highly invasive breast cancer cells. In: BMC Molecular Biology. 2007 ; Vol. 8.
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