Soluble EphB4 inhibition of PDGF-induced RPE migration in vitro.
Soluble EphB4 inhibition of PDGF-induced RPE migration in vitro.
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DOI：
10.1167/iovs.09-3475
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发表时间：
2010-01
影响因子：
4.4
通讯作者：
Hinton DR
中科院分区：
文献类型：
--
作者：
He S;Kumar SR;Zhou P;Krasnoperov V;Ryan SJ;Gill PS;Hinton DR
EphB 4受体（EphB 4）及其配体（EphrinB 2）在调节细胞粘附、生长和迁移中起重要作用。本研究的目的是确定通过可溶性EphB 4（sEphB 4）阻断EphB 4对由血小板衍生生长因子-BB（PDGF）诱导的视网膜色素上皮（RPE）细胞迁移和增殖的影响，并建立其与增殖性玻璃体视网膜病变（PVR）的相关性。通过共聚焦显微镜评估EphB 4和EphrinB 2在早期传代的人RPE细胞和人PVR膜中的表达。分别使用改良的Boyden室测定法和MTT测定法评价sEphB 4（0.1-3 μ g/ml）对PDGF（20 ng/ml）诱导的RPE迁移和增殖的作用。MTT法检测细胞与基底膜基质及纤维连接蛋白的粘附。在暴露于sEphB 4后，通过Western印迹测定RPE中FAK和p42/44 MAP激酶的磷酸化。使用免疫沉淀测定证明sEphB 4对EphB 4/EphrinB 2磷酸化的影响。EphrinB 2和EphB 4在体外在人RPE细胞上和在人PVR膜内的细胞中表达。sEphB 4在体外阻断RPE细胞中EphB 4和EphrinB 2的磷酸化。sEphB 4抑制PDGF刺激的RPE迁移（P<0.01）。sEphB 4以剂量依赖性方式抑制RPE的粘附和增殖（P<0.05）。PDGF诱导的FAK和MAP激酶的磷酸化被sEphB 4抑制。EphB 4和EphrinB 2在RPE细胞和PVR膜中表达。sEphB 4抑制PDGF诱导的RPE细胞粘附、增殖和迁移。这种作用可能是由于抑制FAK和MAP激酶磷酸化引起的。
EphB4 receptor (EphB4) and its ligand (EphrinB2) play an important role in the regulation of cell adhesion, growth and migration. The purpose of this study was to determine the effects of EphB4 blockade by soluble EphB4 (sEphB4) on retinal pigment epithelial (RPE) cell migration and proliferation, induced by platelet-derived growth factor-BB (PDGF), and establish its relevance to proliferative vitreoretinopathy (PVR). The expression of EphB4 and EphrinB2 in early passage human RPE cells and in human PVR membranes was evaluated by confocal microscopy. The effect of sEphB4 (0.1–3 ug/ml) on PDGF (20 ng/ml)-induced RPE migration and proliferation was evaluated using a modified Boyden chamber assay, and MTT assay, respectively. Attachment onto basement membrane matrix and fibronectin was assayed by MTT. Phosphorylation of FAK and p42/44 MAP Kinase in RPE was determined by Western blot after exposure to sEphB4. The effect of sEphB4 on phosphorylation of EphB4/EphrinB2 was demonstrated using immunoprecipitation assays. EphrinB2 and EphB4 were expressed on human RPE cells in vitro, and in cells within human PVR membranes. sEphB4 blocked EphB4 and EphrinB2 phosphorylation in RPE cells in vitro. sEphB4 reduced RPE migration in response to PDGF stimulation (P<0.01). Similarly, sEphB4 inhibited RPE attachment and proliferation in a dose-dependent manner (P<0.05). PDGF-induced phosphorylation of FAK and MAP Kinase was inhibited by sEphB4. EphB4 and EphrinB2 are expressed in RPE cells and PVR membranes. sEphB4 inhibits PDGF induced RPE cell attachment, proliferation and migration. This effect may result from inhibition of FAK and MAP Kinase phosphorylation.
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影响因子：
4.4
作者：
He, SK;Ding, Y;Hinton, DR
通讯作者：
Hinton, DR
影响因子：
50.5
作者：
Alam, S. M.;Fujimoto, J.;Tamaya, T.
通讯作者：
Tamaya, T.
影响因子：
3.7
作者：
Hinton, DR;He, SK;Law, RE
通讯作者：
Law, RE
影响因子：
6
作者：
Nakada, M;Niska, JA;Berens, ME
通讯作者：
Berens, ME
影响因子：
10.5
作者：
Adams, RH;Wilkinson, GA;Klein, R
通讯作者：
Klein, R