CAJ | 학술논문

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.
목적연구가배산납(SCA)유도내피세포조망이급대암세포표체VEGF화Ⅳ형효원매적영향.방법채용류식세포술、DNA응효전영화형태학방법검측ECV304세포조망.Western blotting분석용우관찰암세포VEGF표체.채용매보방법검측Ⅳ형효원매대저물적강해.용ELISA방법검측Ⅳ형효원매화상관단극륭항체적결합.결과SCA정시간의뢰성화제량의뢰성지유도ECV304세포조망.100화250μg·mL-1SCA작용ECV304세포48 h,응효전영현시DNA제대.경SCA작용적ECV304세포형태현시핵내DNA농취,출현조망소체,형광염색검사정현강적람색심염핵.SCA억제VEGF재간암HepG-2세포화전렬선암DU145세포적표체.SCA정제량의뢰성억제폐암PG세포분비적Ⅳ형효원매적강해활성;병억제단항3D6여Ⅳ형효원매적결합.결론 SCA가유도내피세포조망,병억제암세포VEGF표체급Ⅳ형효원매적강해활성.제시SCA가영향종류혈관생성급기미배경.
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 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.