Angiostatin K1-3 induces E-selectin via AP1 and Ets1: A mediator for anti-angiogenic action of K1-3

Y. H. Chen, Y. H. Huang, H. L. Wu, M. P. Wu, W. T. Chang, Y. Z. Kuo, K. C. Lu, Li Wha Wu

Research output: Contribution to journalArticlepeer-review

3 Citations (Scopus)

Abstract

Background: Angiostatin, a circulating angiogenic inhibitor, is an internal fragment of plasminogen and consists of several isoforms, K1-3 included. We previously showed that K1-3 was the most potent angiostatin to induce E-selectin mRNA expression. The purpose of this study was to identify the mechanism responsible for K1-3-induced E-selectin expression and investigate the role of E-selectin in the anti-angiogenic action of K1-3. Methods and results: Quantitative real time RT-PCR and Western blotting analyses confirmed a time-dependent increase of E-selectin mRNA and protein induced by K1-3. Subcellular fractionation and immunofluorescence microscopy showed the co-localization of K1-3-induced E-selectin with caveolin 1 (Cav1) in lipid rafts in which E-selectin may behave as a signaling receptor. Promoter-driven reporter assays and site-directed mutagenesis showed that K1-3 induced E-selectin expression via promoter activation and AP1 and Ets-1 binding sites in the proximal E-selectin promoter were required for E-selectin induction. The in vivo binding of both protein complexes to the proximal promoter was confirmed by chromatin immunoprecipitation (ChIP). Although K1-3 induced the activation of ERK1/2 and JNK, only repression of JNK activation attenuated the induction of E-selectin by K1-3. A modulatory role of E-selectin in the anti-angiogenic action of K1-3 was manifested by both overexpression and knockdown of E-selectin followed by cell proliferation assay. Conclusions: We show that K1-3 induced E-selectin expression via AP1 and Ets-1 binding to the proximal E-selectin promoter (-356/+1), which was positively mediated by JNK activation. Our findings also demonstrate E-selectin as a novel target for the anti-angiogenic therapy.

Original languageEnglish
Pages (from-to)1953-1961
Number of pages9
JournalJournal of Thrombosis and Haemostasis
Volume6
Issue number11
DOIs
Publication statusPublished - 2008 Oct 24

All Science Journal Classification (ASJC) codes

  • Hematology

Fingerprint Dive into the research topics of 'Angiostatin K1-3 induces E-selectin via AP1 and Ets1: A mediator for anti-angiogenic action of K1-3'. Together they form a unique fingerprint.

Cite this