β2-Glycoprotein I inhibits endothelial cell migration through the nuclear factor κB signalling pathway and endothelial nitric oxide synthase activation

Abstract

β₂-GPI (β₂-glycoprotein I) is a plasma glycoprotein ascribed with an anti-angiogenic function; however, the biological role and molecular basis of its action in cell migration remain unknown. The aim of the present study was to assess the contribution of β₂-GPI to HAEC (human aortic endothelial cell) migration and the details of its underlying mechanism. Using wound healing and Boyden chamber assays, we found that β₂-GPI inhibited endothelial cell migration, which was restored by its neutralizing antibody. NF-κB (nuclear factor κB) inhibitors and lentiviral siRNA (small interfering RNA) silencing of NF-κB significantly attenuated the inhibitory effect of β₂-GPI on cell migration. Moreover, β₂-GPI was found to induce IκBα (inhibitor of NF-κB) phosphorylation and translocation of p65 and p50. We further demonstrated that mRNA and protein levels of eNOS [endothelial NO (nitric oxide) synthase] and NO production were all increased by β₂-GPI and these effects were remarkably inhibited by NF-κB inhibitors and siRNAs of p65 and p50. Furthermore, β₂-GPI-mediated inhibition of cell migration was reversed by eNOS inhibitors and eNOS siRNAs. The findings of the present study provide novel insight into the ability of β₂-GPI to inhibit endothelial cell migration predominantly through the NF-κB/eNOS/NO signalling pathway, which indicates a potential direction for clinical therapy in vascular diseases.

Reduced beta2-glycoprotein I protects macrophages from ox-LDL-induced foam cell formation and cell apoptosis

“…Results showed a statistical significance using the concentration of 100 μmmol/L beta2-GPI or reduced beta2-GPI. Synthesizes related references [13,14,18,19] and our preliminary experiment, the concentration of 100 μmmol/L was selected for the mechanism study.…”

Binding of plasma proteins to titanium dioxide nanotubes with different diameters

“…30,32 SAA is a relatively small protein of ~12 kDa 33 with ~80% alpha helical conformation. 34 β2GPI, also known as apolipoprotein H, 35 is a multifunctional 53 kDa glycosylated plasma protein 36 with physiological values of 20-300 mg/L. 37 The protein consists of 326 amino acids folded into five domains, where the fifth domain is positively charged, with a lysine-rich region 281 CKNKEKKC, 288 a hydrophobic loop 313 LAFW, 316 and extended C-terminal domain, that enable binding to negatively charged phospholipids.…”

Structural and functional characterization of β2‐glycoprotein I domain 1 in anti‐melanoma cell migration

“…[12][13][14][15] We showed previously that purified β 2 -GPI inhibits cell migration, proliferation, and angiogenesis in human aortic endothelial cells. [9][10][11] However, the role of natural β 2 -GPI in tumor development remains unknown. The present study showed that β 2 -GPI is a negative regulator of cell migration, proliferation, and invasion in melanoma cells.…”

“…We previously showed that β 2 -GPI plays an essential role in the suppression of vascular endothelial cell growth, cell migration, and angiogenesis under the regulation of nuclear factor-kappa B (NF-кB) signaling and vascular endothelial growth factor (VEGF)-mediated phosphorylation of VEGFR2, ERK1/2 and Akt pathways. [9][10][11] It is well established that abnormal cell migration and proliferation are closely related to carcinogenesis. [12][13][14][15] Furthermore, many studies have reported that inhibition of cell migration decreases tumor growth in multiple cancers.…”

“…2.2 | Purification of β 2 -GPI and recombinant β 2 -GPI polypeptides β 2 -Glycoprotein I was purified from human plasma using an established protocol. 9 Briefly, plasma β 2 -GPI was isolated by 3% perchloric acid precipitation followed by heparin-sepharose affinity chromatography (HiTrap Heparin; GE Healthcare Bio-Sciences, Marlborough, MA, USA). Purity of β 2 -GPI was confirmed by SDS-PAGE and western blot analysis.…”