Gastrin enhances the anglogenic potential of endothelial cells via modulation of heparin-binding epidermal-like growth factor
Gastrin enhances the anglogenic potential of endothelial cells via modulation of heparin-binding epidermal-like growth factor
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DOI：
10.1158/0008-5472.can-05-0280
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发表时间：
2006-04-01
期刊：
影响因子：
11.2
通讯作者：
Watson, SA
中科院分区：
文献类型：
--
作者：
Clarke, PA;Dickson, JH;Watson, SA
本研究检验了胃泌素是否通过肝素结合表皮生长因子样生长因子 (HB-EGF) 的表达来调节内皮细胞活性。在酰胺化胃泌素 17 (G17) 和甘氨酸延伸胃泌素 17 (GlyG17) 肽存在下评估人脐血管内皮细胞 (HUVEC) 的小管形成。通过定量逆转录 PCR、免疫细胞化学和蛋白质印迹法测量 HB-EGF 基因和蛋白表达，并通过 ELISA 测量 HB-EGF 脱落。通过蛋白质印迹法评估基质金属蛋白酶 MMP-2、MMP-3 和 MMP-9。鸡绒毛尿囊膜研究测量了胃泌素的体内血管生成潜力，并在高胃泌素血症 APC(Min) 小鼠的大肠癌前病变中评估了微血管密度 (MVD)。还在人类结直肠肿瘤和正常切缘中检查了 MVD，并与血清酰胺化胃泌素水平（通过 RIA）和 HB-EGF 蛋白表达（通过免疫组织化学）相关。 HUVEC 细胞响应 G17 (186%, P < 0.0005) 和 GlyG17 (194%, P < 0.0005) 显示增加的肾小管和结节形成。该作用可被胆囊收缩素-2 受体 (CCK-2R) 拮抗剂 JB95008 和 JMV1155 以及胃泌素和 HB-EGF 抗血清阻断。胃泌素肽增加 HUVEC 和微血管源性内皮细胞中 HB-EGF 基因表达/蛋白质分泌以及 MMP-2、MMP-3 和 MMP-9 的水平。在绒毛尿囊膜测定中，G17 促进血管生成，并且高胃泌素血症 APC(Min) 小鼠的癌前大肠组织中的 MVD 显着升高。从临床情况来看，结直肠腺癌周围正常粘膜的MVD与患者血清胃泌素水平和HB-EGF表达相关。胃泌素肽通过 CCK-2R 发挥作用，增强血管生成模型中的内皮细胞活性。这可能是通过增强 HB-EGF 的表达和脱落来介导的，这可能是由于基质金属蛋白酶活性的增加所致。这种促血管生成作用转化为体内和人类情况，并可能增加恶性肿瘤中胃泌素肽的致瘤特性。
This study examined whether gastrin modulates endothelial cell activity via heparin-binding epidermal growth factor-like growth factor (HB-EGF) expression. Human umbilical vascular endothelial cells (HUVEC) were assessed for tubule formation in the presence of amidated gastrin-17 (G17) and glycine-extended gastrin-17 (GlyG17) peptides. HB-EGF gene and protein expressions were measured by quantitative reverse transcription-PCR, immunocytochemistry, and Western blotting, and HB-EGF shedding by ELISA. Matrix metalloproteinases MMP-2, MMP-3, and MMP-9 were assessed by Western blotting. Chick chorioallantoic membrane studies measured the in vivo angiogenic potential of gastrin and microvessel density (MVD) was assessed in large intestinal premalignant lesions of hypergastrinaemic APC(Min) mice. MVD was also examined in human colorectal tumor and resection margin normals and correlated with serum-amidated gastrin levels (via RIA) and HB-EGF protein expression (via immunohistochemistry). HUVEC cells showed increased tubule and node formation in response to G17 (186%, P < 0.0005) and GlyG17 (194%, P < 0.0005). This was blockaded by the cholecystokinin-2 receptor (CCK-2R) antagonists JB95008 and JMV1155 and by antiserum to gastrin and HB-EGF. Gastrin peptides increased HB-EGF gene expression/protein secretion in HUVEC and microvessel-derived endothelial cells and the levels of MMP-2, MMP-3, and MMP-9. G17 promoted angiogenesis in a chorioallantoic membrane assay, and MVD was significantly elevated in premalignant large intestinal tissue from hypergastrinaemic APC(Min) mice. lit terms of the clinical situation, MVD in the normal mucosa surrounding colorectal adenocarcinomas correlated with patient serum gastrin levels and HB-EGF expression. Gastrin peptides, acting through the CCK-2R, enhance endothelial cell activity in models of angiogenesis. This may be mediated through enhanced expression and shedding of HB-EGF, possibly resulting from increased activity of matrix metalloproteinases. This proangiogenic effect translates to the in vivo and human situations and may add to the tumorigenic properties attributable to gastrin peptides in malignancy.