摘要
目的研究胃泌素对人脐静脉血管内皮细胞（HUVEC）体外增殖和管状结构形成的作用。方法免疫细胞化学等检测HUVEC中胃泌素受体（CCK—BR）的表达；用10和100nmoL胃泌素处理HUVEC后，MTT和软琼脂克隆形成实验检测细胞增殖能力，小管形成实验观察HUVEC管状结构生成情况；实时荧光定量PCR和酶联免疫吸附法检测HUVEC中血管内皮生长因子（VEGF）和低氧诱导因子．1a（HIF，la）基因的表达。结果HUVEC表达CCK—BR，经10和100nmoL胃泌素处理后．细胞增殖率和克隆形成率显著增加（P〈0．01），细胞中VEGF和HIF-1a基因的表达量高于对照组（P〈0．01），HUVEC形成的半环状和环状管状结构数也高于对照组（P〈0．01）。结论胃泌素与HUVEC表达的CCK．BR结合后，上调VEGF和HIF．1a基因的表达，促进HUVEC体外增殖及管状结构的形成能力。
Objective To study the effect of gastrin on the proliferation and angiogenesis of human umbilical vascular endothelial （HUVE） cell in vitro. Methods The immunocytochemistry assay, realtime-PCR, and Western blot were used to detect the gastrin receptor （CCK-BR） expression in HUVE cells. After HUVE cells were treated with 10 and 100 nmol/L gastrin for 72 h, MTT and soft agar colony formation assay were used to test the cell proliferation rate and colony formation rate, and the vascular-like structures were observed by three-dimensional culture of HUVE cells. The half-ring and ring vascular numbers were counted with five random visions. The vascular endothelial growth factor （VEGF） and hypoxia-inducible factor-la （HIF-la） mRNA and protein levels in HUVE cells were detected by realtime PCR and ELISA assay respectively. Results HUVE cells expressed CCK-BR. After the treatment with 10 and 100 nmol/L gastrin, the cell proliferation rate was increased by 48.48% and 82.82 % compared to the control group, and colony formation rate was increased to 31.33% and 45.67 % as compared with 15.33% in the control group（ P〈0. O1 ）. Relative expression quantities of VEGF and HIF-la genes were 2.3 and 4.6 folds （VEGF） and 20.76 and 26.77 folds （ HIF-1 a） than those in the control group. The concentration of VEGF protein in culture medium was 22l and 392 Ixg/mg protein higher than that in control group. The numbers of half-ring and ring vascular structures were （ 14. 00 ± 3.130, 39.33 ±7.57 and 34.33 ±4.50 ）/vision and （ 8.33 ± 2.51,41.33 ±5.85 and 37.67 ±3.51 ）/vision in control, 10 and 100 nmol/L gastrin-treated groups, respectively （ P〈O. O1 ）. Conclusion Gastrin up-regulates the expression of VEGF and HIF-1 a genes in HUVE cells and promotes cell proliferation and vascular-like structure formation of HUVE cells in vitro by being combined to CCK-BR, which may be involved in the development and metastasis of gastric cancer.
出处
《中华内分泌代谢杂志》
CAS
CSCD
北大核心
2013年第7期598-602,共5页
Chinese Journal of Endocrinology and Metabolism
基金
国家自然科学基金项目（31060122）
贵州省科技攻关计划基金（黔科合sY字[2011]3067号）
贵州省科学技术基金（黔科合J字[2012]2039号）
