瑶药消瘤藤体外抗肿瘤活性部位筛选

Screening of the Anti Tumor Extracts in Pileostegia Tomentella Hand. Mazz(Xiaoliuteng)

目的:筛选瑶药消瘤藤中的体外抗肿瘤部位。方法:采用溶剂提取,硅胶柱色谱法对消瘤藤中的总香豆素及总苷类成分进行提取分离,紫外分光光度法测定总香豆素和总苷的含量;CCK8法和克隆形成实验检测对肿瘤细胞的增殖活力的影响、划痕实验及Transwell侵袭小室观察肿瘤细胞迁移、侵袭能力的改变,蛋白免疫印迹法(Western blot)检测Bcl-2、Caspase3和NF-κB的蛋白表达。结果:消瘤藤总香豆素中香豆素含量为65.43%,总苷含量为52.02%。消瘤藤总香豆素部位可呈剂量依赖方式显著提高对人肝癌HepG2细胞、人结肠癌HT-29细胞、人神经胶质瘤U87细胞及人乳腺癌MDA-MB-231细胞增殖的抑制率,显著抑制HT-29细胞的克隆、迁移及侵袭能力,显著下调Bcl-2、NF-κB的蛋白表达,上调Caspase3的蛋白表达。结论:总香豆素部位为消瘤藤抗肿瘤的活性部位,能抑制HT-29细胞的增殖、克隆、迁移及侵袭能力,其机制可能与上调Caspase3蛋白表达,下调Bcl-2和NF-κB蛋白表达有关。

Objective:To screen the anti-tumor active extracts from Pileostegia tomentella Hand.Mazz(Xiaoliuteng).Methods:Solvent extraction method and silica column chromatography were used to isolate total coumarins and total glycosides in Xiaoliuteng,their contents were determined by ultraviolet spectrophotometry.CCK8 method and Colony formation assay were used to detect the proliferation ability of the cells.Transwell assay and scratch test were used to explore the changes of invasion and migration of tumors cells.Western blot was performed to detect the protein expressions of Bcl-2,Caspase3 and NF-κB.Results:The content of coumarin was 65.43%,the content of total glycosides was 52.02%.The total coumarins significantly increased the inhibition rate on cell lines HepG2,HT-29,U87 and MDA-MB-231.It also significantly inhibited the clone,migration and invasion of HT-29 cells,and down regulated the protein expressions of Bcl-2 and NF-κB,up regulated the expression of Caspase3.Conclusion:Total coumarins is the active antitumor component from Pileostegia tomentella Hand.Mazz(Xiaoliuteng),it can inhibit the proliferation,clone migration and invasion of HT-29 cells,which may be related to up-regulating Caspase3 protein and down-regulating the protein expressions of Bcl-2 and NF-κB.