肝细胞生长因子及受体促进人大肠癌细胞血管内皮生长因子表达

Up-regualtion of vascular endothelial growth factor by hepatocyte growth factor in human colorectal carcinoma cells

目的:观察不同浓度肝细胞生长因子对体外培养的大肠癌细胞血管内皮生长因子(VEGF)表达的影响。方法:体外培养人大肠癌细胞,分别加入不同浓度的人重组肝细胞生长因子和或其受体抑制剂Herbimycin A(HA),用Western blot、RT-PCR、ELISA等方法检测肝细胞生长因子对其受体磷酸化表达及大肠癌细胞VEGF表达的影响。结果:肝细胞生长因子在10～100ng/ml浓度范围内较对照组相比显著促进大肠癌细胞VEGF mRNA(增加了4～5倍)、蛋白质表达(增加了3～10倍);Western blot检测肝细胞生长因子引起其受体磷酸化,并与其诱导的VEGF表达相关。结论:体外培养条件下,肝细胞生长因子呈剂量依赖性促进大肠癌细胞VEGF表达,其表达与其受体磷酸化水平相关;其受体磷酸化抑制剂可以抑制大肠癌细胞VEGF表达。

Objective：To observe the effect of human recombinant hepatocyte growth factor （HGF） on the expression of vascular endothelial growth factor（VEGF） in colorectal carcinoma（CRC） cells in vitro. Methods：Human CRC cell lines SW480, LOVO were cultured with different concentrations of HGF. The culture supematants were collected to detect the production of VEGF by ELISA assay. The effect of HGF on phosphorylation of its receptor c-Met was detected by Western blot assay. The expression of VEGF mRNA stimulated by HGF was investigated by reverse transcription PCR（RT-PCR）. Resuits：HGF increased the expression of VEGF （mRNA 4-5-fold and protein 3-10-fold） in CRC cell lines; HGF induced the phosphorylation of its receptor c-Met in a dose-dependent manner. Conclusion：In vitro, HGF can up-regulate the expression of VEGF in CRC cells, which correlates with the phosphorylation of its c-Met induced by HGF. Inhibitor of phosphorylation-c-Met, HA, can inhibit the expression of VEGF in CRC cells induced by HGF.