The Heparin Binding Motif of Endostatin Mediates Its Interaction with Receptor Nucleolin

Fu, Yan; Chen, Yang; Luo, Xi; Liang, Yun; Shi, Hubing; Gao, Lei; Zhan, Shunli; Zhou, Daifu; Luo, Yongzhang

doi:10.1021/bi901265z

Cited by 20 publications

(21 citation statements)

References 27 publications

Supporting

Mentioning

Contrasting

Order By: Relevance

“…Nucleolin is a cell surface receptor in angiogenic EC cells [48] and shuttles between the nucleus, cytoplasm and cell surface. Studies have shown that anti-angiogenic endostatin was taken up by the cells and transported into the nuclei of EC via nucleolin receptor and subsequently prevented growth and angiogenesis [49,50]. Therefore, the miR200-b DNA NP might be internalized and shuttled to the nucleus of EC cells via nucleolin receptors, subsequently expresses miR200-b that consequently knocks down VEGFR-2 expression as shown by both messenger and protein analyses (Fig.…”

Section: Discussionmentioning

confidence: 98%

Nanoparticle-mediated miR200-b delivery for the treatment of diabetic retinopathy

Mitra

Nichols

Guo

et al. 2016

Journal of Controlled Release

View full text Add to dashboard Cite

We recently reported that the Ins2Akita mouse is a good model for late-onset diabetic retinopathy. Here, we investigated the effect of miR200-b, a potential anti-angiogenic factor, on VEGF receptor 2 (VEGFR-2) expression and to determine the underlying angiogenic response in mouse endothelial cells, and in retinas from aged Ins2Akita mice. MiR200-b and its native flanking sequences were amplified and cloned into a pCAG-eGFP vector directed by the ubiquitous CAG promoter (namely pCAG-miR200-b-IRES-eGFP). The plasmid was compacted by CK30PEG10K into DNA nanoparticles (NPs) for in vivo delivery. Murine endothelial cell line, SVEC4-10, was first transfected with the plasmid. The mRNA levels of VEGF and VEGFR-2 were quantified by qRT-PCR and showed significant reduction in message expression compared with lipofectamine-transfected cells. Transfection of miR200-b suppressed the migration of SVEC4-10 cells. There was a significant inverse correlation between the level of expression of miR200-b and VEGFR-2. Intravitreal injection of miR200-b DNA NPs significantly reduced protein levels of VEGFR-2 as revealed by western blot and markedly suppressed angiogenesis as evaluated by fundus imaging in aged Ins2Akita mice even after 3 months of post-injection. These findings suggest that NP-mediated miR200-b delivery has negatively regulated VEGFR-2 expression in vivo.

show abstract

Section: Discussionmentioning

confidence: 98%

Nanoparticle-mediated miR200-b delivery for the treatment of diabetic retinopathy

Mitra

Nichols

Guo

et al. 2016

Journal of Controlled Release

View full text Add to dashboard Cite

show abstract

“…However, the mechanisms by which heparin and thrombin mediate their effects on angiogenesis remain uncertain. Previous work suggests that the antiangiogenic effects of heparin are unrelated to its anticoagulant actions (37,44,45). However, this is still in dispute.…”

Section: Discussionmentioning

confidence: 99%

Heparin Impairs Angiogenesis through Inhibition of MicroRNA-10b

Shen

Fang

et al. 2011

Journal of Biological Chemistry

View full text Add to dashboard Cite

Heparin, which has been used as an anticoagulant drug for decades, inhibits angiogenesis, whereas thrombin promotes tumor-associated angiogenesis. However, the mechanisms underlying the regulation of angiogenesis by heparin and thrombin are not well understood. Here, we show that microRNA-10b (miR-10b) is down-regulated by heparin and up-regulated by thrombin in human microvascular endothelial cells (HMEC-1). Overexpression of miR-10b induces HMEC-1 cell migration, tube formation, and angiogenesis, and downregulates homeobox D10 (HoxD10) expression via direct binding of miR-10b to the putative 3 UTR of HoxD10. In addition, HMEC-1 cell migration and tube formation are induced by HoxD10 knockdown, whereas angiogenesis is arrested when HoxD10 expression is increased after anti-miR-10b or heparin treatments. Furthermore, expression of miR-10b and its transcription factor Twist are up-regulated by thrombin, whereas HoxD10 expression is impaired by thrombin. Using quartz crystal microbalance analysis, we show that heparin binds to thrombin, thereby inhibiting thrombin-induced expression of Twist and miR-10b. However, the expression of miR-10b is not attenuated by heparin any more after thrombin expression is silenced by its siRNA. Interestingly, we find that heparin attenuates miR10b expression and induces HoxD10 expression in vivo to inhibit angiogenesis and impair the growth of MDA-MB-231 tumor xenografts. These results provide insight into the molecular mechanism by which heparin and thrombin regulate angiogenesis.Thrombosis is considered an early clinical indication and frequent complication of cancer (1, 2). Malignant tumors often exhibit increased expression of tissue factor and cancer procoagulant, which can be followed by activation of cell surface protease receptors and fibrin generation (3). In addition, tumor cells can interact with blood cells, particularly monocytes, macrophages, and platelets, leading to the generation of thrombin and thrombosis through the clotting cascade or platelet activation (4). Moreover, aggressive antitumor therapies such as chemotherapy, radiation, and surgery also increase the risk of thrombosis.Positive feedback signaling loops exist between tumor tissue and the coagulation system (5, 6). For instance, it is widely accepted that elements of the coagulation and fibrinolytic system may aid in cancer cell survival, proliferation, invasion, and metastasis, as well as tumor angiogenesis (7). Therefore, inhibition of the activation of coagulation could be a useful antitumor strategy. A number of studies have shown that anticoagulant drugs can extend survival in patients with certain types of cancer (8), and studies are currently ongoing to confirm the effects of anticoagulant therapy in a range of tumor types. However, the molecular mechanisms involved in the action of anticoagulants in this process are not well understood.Heparins, in particular those of low molecular weight, are effective in the prevention and treatment of thromboembolic events in cancer patients (9, 10). As early...

show abstract

“…Nucleolin was reported to function as cell surface receptor for molecules like midkine or pleiotrophin [59-61]. Interestingly, as reported for HDGF nucleolin can also bind to certain glycosaminoglycans [3,8,42,62,63] and heparin binding motifs of potential ligands can function as binding motifs for nucleolin [64]. Similar to ribonucleic acids GAGs have been demonstrated to support a wide variety of protein-protein interactions.…”

Section: Discussionmentioning

confidence: 99%

Hepatoma-derived growth factor and nucleolin exist in the same ribonucleoprotein complex

et al. 2013

View full text Add to dashboard Cite

BackgroundHepatoma-derived growth factor (HDGF) is a protein which is highly expressed in a variety of tumours. HDGF has mitogenic, angiogenic, neurotrophic and antiapoptotic activity but the molecular mechanisms by which it exerts these activities are largely unknown nor has its biological function in tumours been elucidated. Mass spectrometry was performed to analyse the HDGFStrep-tag interactome. By Pull–down-experiments using different protein and nucleic acid constructs the interaction of HDGF and nucleolin was investigated further.ResultsA number of HDGFStrep-tag copurifying proteins were identified which interact with RNA or are involved in the cellular DNA repair machinery. The most abundant protein, however, copurifying with HDGF in this approach was nucleolin. Therefore we focus on the characterization of the interaction of HDGF and nucleolin in this study. We show that expression of a cytosolic variant of HDGF causes a redistribution of nucleolin into the cytoplasm. Furthermore, formation of HDGF/nucleolin complexes depends on bcl-2 mRNA. Overexpression of full length bcl-2 mRNA increases the number of HDGF/nucleolin complexes whereas expression of only the bcl-2 coding sequence abolishes interaction completely. Further examination reveals that the coding sequence of bcl-2 mRNA together with either the 5′ or 3′ UTR is sufficient for formation of HDGF/nucleolin complexes. When bcl-2 coding sequence within the full length cDNA is replaced by a sequence coding for secretory alkaline phosphatase complex formation is not enhanced.ConclusionThe results provide evidence for the existence of HDGF and nucleolin containing nucleoprotein complexes which formation depends on the presence of specific mRNAs. The nature of these RNAs and other components of the complexes should be investigated in future.

show abstract

The Heparin Binding Motif of Endostatin Mediates Its Interaction with Receptor Nucleolin

Cited by 20 publications

References 27 publications

Nanoparticle-mediated miR200-b delivery for the treatment of diabetic retinopathy

Nanoparticle-mediated miR200-b delivery for the treatment of diabetic retinopathy

Heparin Impairs Angiogenesis through Inhibition of MicroRNA-10b

Hepatoma-derived growth factor and nucleolin exist in the same ribonucleoprotein complex

Contact Info

Product

Resources

About