咖啡酸钠诱导内皮细胞凋亡及抑制癌细胞VEGF的表达(英文)  

Sodium caffeate induces endothelial cell apoptosis and inhibits VEGF expression in cancer cells

作　　者：徐峰[1] 欧阳志钢[1] 张胜华[1] 宋丹青[1] 邵荣光[1] 甄永苏[1] 

机构地区：[1]中国医学科学院,中国协和医科大学医药生物技术研究所,北京100050

出　　处：《药学学报》2006年第6期572-576,共5页Acta Pharmaceutica Sinica

基　　金：Key Basic Research Development Program Foundation of China (2002CB513108).

摘　　要：目的研究咖啡酸钠(SCA)诱导内皮细胞凋亡以及对癌细胞表达VEGF和IV型胶原酶的影响。方法采用流式细胞术、DNA凝胶电泳和形态学方法检测ECV304细胞凋亡。Western blotting分析用于观察癌细胞VEGF表达。采用酶谱方法检测IV型胶原酶对底物的降解。用ELISA方法检测IV型胶原酶和相关单克隆抗体的结合。结果 SCA呈时间依赖性和剂量依赖性地诱导ECV304细胞凋亡。100和250μg·mL-1SCA作用ECV304细胞48 h,凝胶电泳显示DNA梯带。经SCA作用的ECV304细胞形态显示核内DNA浓聚,出现凋亡小体,荧光染色检查呈现强的蓝色深染核。SCA抑制VEGF在肝癌HepG-2细胞和前列腺癌DU145细胞的表达。SCA呈剂量依赖性抑制肺癌PG细胞分泌的IV型胶原酶的降解活性;并抑制单抗3D6与IV型胶原酶的结合。结论 SCA可诱导内皮细胞凋亡,并抑制癌细胞VEGF表达及IV型胶原酶的降解活性。提示SCA可影响肿瘤血管生成及其微环境。Aim To investigate the induction of endothelial cell apoptosis and the suppression of VEGF expression in cancer cells by sodium caffeate （SCA）. Methods Apoptosis of transformed human umbilical vein endothelial cells （ECV304 cell line） was detected by flow cytometry, DNA electrophoresis assay and morphological assessment. Western blotting analysis was applied for determination of VEGF expression in cancer cells. Substrate degradation by type IV collagenase was measured by zymography. ELISA was used to detect the binding of type IV collagenase with relevant monoclonal antibody. Results SCA induced ECV304 cell apoptosis in a time- and dose-dependent manner. After treatment with 100 and 250 μg·mL^-1 of SCA for 48 h, DNA laddering appeared. SCA treated cells showed strong blue fluorescence and distinct changes of nuclear morphology, such as pyknosis and the occurrence of apoptotic bodies. VEGF expression in hepatoma HepG-2 cells and prostate carcinoma DU145 cells was reduced after SCA treatment. The degradation activity of type IV collagenase including MMP-2 and MMP-9 secreted by giant cell pulmonary carcinoma PG cells was inhibited by SCA in a dose-dependent manner. SCA also reduced the binding of mAb 3D6, a relevant monoclonal antibody, to type IV collagenase. Conclusion SCA can induce endothelial cell apoptosis and inhibit VEGF expression as well as type IV collagenase activity in cancer cells. SCA might be active in modulating tumor angiogenesis and the microenvironment.

关 键 词：细胞凋亡 内皮细胞 微环境 咖啡酸钠 VEGF 

分 类 号：R963[医药卫生—微生物与生化药学]