摘要
目的:研究知母皂苷B-Ⅱ(TB-Ⅱ)抑制人肺癌A549细胞增殖和迁移的机制。方法:使用0(空白对照)、1、10、100μg/ml的TB-Ⅱ处理A549细胞48 h后,分别提取细胞总RNA及总蛋白,采用实时定量荧光-聚合酶链反应和Western blot法分别检测细胞内白细胞介素18(IL-18)m RNA及蛋白表达;通过转染沉默A549细胞内IL-18基因,比较未转染组、阴性对照组和转染组细胞内IL-18 m RNA及蛋白表达,然后用10μg/ml的TB-Ⅱ处理转染后的细胞24、48、72 h,采用CCK-8法检测细胞增殖活性,划线法观察细胞迁移能力变化。结果:与空白对照比较,TB-Ⅱ处理后A549细胞内IL-18 m RNA及蛋白表达均增加(P<0.05或P<0.01),且与浓度呈正相关。与未转染组比较,转染组细胞内IL-18 m RNA及蛋白表达均降低(P<0.01);与TB-Ⅱ处理的未转染细胞比较,TB-Ⅱ处理72 h后的转染细胞增殖活性和迁移能力均增强(P<0.01)。结论:TB-Ⅱ可通过上调IL-18基因的表达来抑制A549细胞的增殖和迁移。
OBJECTIVE：To study the inhibitory mechanism of timosaponin B-Ⅱ（TB-Ⅱ）on the proliferation and migration of human lung cancer A549 cells. METHODS：A549 cells were treated with TB-Ⅱ [0（blank control）,1,10 and 100 μg/ml] for48 h,and total RNA and total protein were extracted respectively. Real time fluorescence quantitative-PCR and Western blot were used to detect m RNA and protein levels of IL-18. IL-18 in A549 cells was silenced by transfection;the expression of IL-18 m RNA and protein were compared among untransfection group,negative control group and transfection group;and then human lung cancer A549 cells with silenced gene were treated with 10 μg/ml TB-Ⅱ for 24,48 and 72 h. The activity of cell proliferation was detected with CCK-8,and the change of cell migration ability was observed by streak method. RESULTS：Compared with blank control,the expression of IL-18 m RNA and protein in A549 cells all increased after treated with TB- Ⅱ（P0.05 or P0.01）,and were positively correlated with concentration. Compared with untransfection group,the expression of IL-18 m RNA and protein decreased in transfection group（P0.01）. Compared with untransfected cell treated with TB-Ⅱ,the viability and migration ability of A549 cells with transfection gene increased after treated with TB-Ⅱ for 72 h（P0.01）. CONCLUSIONS：TB-Ⅱ can inhibit the proliferation and migration of A549 cells by up-regulating IL-18 gene expression.
出处
《中国药房》
CAS
北大核心
2016年第10期1346-1349,共4页
China Pharmacy
