Gastrin promotes the proliferation and angiogenesis of human umbilical vascular endothelial cell / 中华内分泌代谢杂志

ABSTRACT

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-1a (HIF-1a) 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＜0.01).Relative expression quantities of VEGF and HIF-1a 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 221 and 392 μg/mg protein higher than that in control group.The numbers of half-ring and ring vascular structures were (14.00 ± 3.00,39.33 ± 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＜0.01).Conclusion Gastrin up-regulates the expression of VEGF and HIF-1a 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.