Lefranc Florence, Mijatovic Tatjana, Mathieu Véronique, Rorive Sandrine, Decaestecker Christine, Debeir Olivier, Brotchi Jacques, Van Ham Philippe, Salmon Isabelle, Kiss Robert
Department of Neurosurgery, Erasmus University Hospital.
Clin Cancer Res. 2004 Dec 15;10(24):8250-65. doi: 10.1158/1078-0432.CCR-04-0343.
This study aims to investigate the role of gastrin-17 (G17) on angiogenesis features in gliomas both in vitro and in vivo.
The influences of G17 and G17 receptor antagonists were characterized in vitro in terms of angiogenesis on human umbilical vein endothelial cell (HUVEC) tubulogenesis processes on Matrigel and in vivo with respect to U373 orthotopic glioma xenografts. The influence of phosphatidylinositol 3'-kinase, protein kinase C, and nuclear factor-kappaB inhibitors was characterized in vitro on G17-mediated HUVEC tubulogenesis. G17-mediated release of interleukin (IL)-8 from HUVECs and G17-induced modifications in nuclear factor-kappaB DNA binding activity were characterized by means of specific enzyme-linked immunosorbent assays. The influence of G17 on E- and P-selectin expression was determined by means of computer-assisted microscopy, whereas the influence of E- and P-selectin on HUVEC migration was approached by means of antisense oligonucleotides. The chemotactic influence of G17 and IL-8 on HUVEC migration was characterized by means of computer-assisted videomicroscopy with Dunn chambers.
Messenger RNAs for cholecystokinin (CCK)A, CCKB, and CCKC receptors were present in HUVECs and microvessels dissected from a human glioblastoma. Whereas G17 significantly increased the levels of angiogenesis in vivo in the U373 experimental glioma model and in vitro in the HUVECs, the CCKB receptor antagonist L365,260 significantly counteracted the G17-mediated proangiogenic effects. G17 chemoattracted HUVECs, whereas IL-8 failed to do so. IL-8 receptor alpha (CXCR1) and IL-8 receptor beta (CXCR2) mRNAs were not detected in these endothelial cells. Gastrin significantly (but only transiently) decreased the level of expression of E-selectin, but not P-selectin, whereas IL-8 increased the expression of E-selectin. Specific antisense oligonucleotides against E- and P-selectin significantly decreased HUVEC tubulogenesis processes in vitro on Matrigel.
The present study shows that gastrin has marked proangiogenic effects in vivo on experimental gliomas and in vitro on HUVECs. This effect depends in part on the level of E-selectin activation, but not on IL-8 expression/release by HUVECs.
本研究旨在探讨胃泌素 - 17（G17）在体外和体内对胶质瘤血管生成特征的作用。
在体外，通过基质胶上人脐静脉内皮细胞（HUVEC）成管过程研究G17和G17受体拮抗剂对血管生成的影响；在体内，以U373原位胶质瘤异种移植模型进行研究。在体外，通过磷脂酰肌醇3'-激酶、蛋白激酶C和核因子 - κB抑制剂研究它们对G17介导的HUVEC成管的影响。通过特异性酶联免疫吸附测定法研究G17介导的HUVEC白细胞介素（IL）-8释放及G17诱导的核因子 - κB DNA结合活性变化。通过计算机辅助显微镜测定G17对E - 选择素和P - 选择素表达的影响，而通过反义寡核苷酸研究E - 选择素和P - 选择素对HUVEC迁移的影响。通过带邓恩小室的计算机辅助视频显微镜测定G17和IL - 8对HUVEC迁移的趋化作用。
胆囊收缩素（CCK）A、CCK B和CCK C受体的信使核糖核酸存在于人脐静脉内皮细胞及从人胶质母细胞瘤分离的微血管中。在U373实验性胶质瘤模型中，G17在体内显著增加血管生成水平，在体外对人脐静脉内皮细胞也有同样作用；而CCK B受体拮抗剂L365,260显著抵消G17介导的促血管生成作用。G17对人脐静脉内皮细胞有趋化作用，而IL - 8则无此作用。在这些内皮细胞中未检测到IL - 8受体α（CXCR1）和IL - 8受体β（CXCR2）的信使核糖核酸。胃泌素显著（但只是短暂地）降低E - 选择素的表达水平，但不影响P - 选择素，而IL - 8增加E - 选择素表达。针对E - 选择素和P - 选择素的特异性反义寡核苷酸在体外显著降低基质胶上的人脐静脉内皮细胞成管过程。
本研究表明，胃泌素在体内对实验性胶质瘤及在体外对人脐静脉内皮细胞有显著的促血管生成作用。这种作用部分取决于E - 选择素的激活水平，而不依赖于人脐静脉内皮细胞IL - 8的表达/释放。
