摘要
探讨CPUY013对体外培养人胰腺癌细胞Capan2生长抑制及诱导凋亡作用.采用MTT法、克隆原形成法检测CPUY013对Capan2细胞生长的抑制作用,采用流式细胞仪分析CPUY013对细胞周期的影响,Western Blot法检测TopoⅠ、野生型p53、caspase-3、bcl-2和bax蛋白表达的变化.MTT法、集落形成试验结果显示,CPUY013对体外培养的人胰腺癌细胞Capan2有明显的生长抑制作用,处理72 h的IC50值为7.3×10-7mol/L(MTT法),CPUY013在8.0×10-8mol/L浓度可以明显抑制Ca-pan2细胞的集落形成,抑制率为69.6%.同时,CPUY013可剂量依赖地降低Capan2细胞G1期细胞的比例,升高S期细胞的比例,呈现明显的S期阻滞.CPUY013可下调TopoⅠ蛋白的表达,上调细胞中p53、caspase-3、bax蛋白表达,下调bcl-2蛋白表达,并呈剂量依赖性.CPUY013对Capan2细胞具有明显的生长抑制作用,阻滞细胞周期进程,诱导Capan2细胞凋亡,其可能与降低细胞内TopoⅠ蛋白表达,增加p53、caspase-3、bax蛋白表达,降低bcl-2蛋白表达有关.
To explore the inhibition of CPUY013 ,a novel TOPO Ⅰ inhibitor ,on the proliferation and induced apoptosis in pancreatic cancer cells Capan2 and possible mechanism in vitro. The inhibition of CPUY013 in Capan2 cells is investigated with MTT assay and colony formation assay. Changes of cell cycle are studied by flow cytometry analysis. Western Blot assay is used to detect the protein expression levels of TOPO Ⅰ ,widetype p53, caspase - 3, bcl -2 and bax. The results of MT and colony formation assay showed that CPUY013 had a marked growth inhibition in Capan2 cells and its IC50 is about 7.3×10^-7 mol/L after treatment for 72 h by CPUY013, inhibitory rate on colony formation is about 69.6% at 8.0 ×10^-8 mol/L CPUY013. Cell cycle changes are analyzed by flow cytometry with cells at phase significantly less than those of control and cells at S phase significantly more than those of control . It suggests that CPUY013 could induced apoptosis of Capan2 ceils with the cell cycle arrest on S phase in distinct dose-dependent manner. The expressions of TOPO Ⅰ ,p53, bax, caspase-3 protein increased and the expression of bcl-2 protein is decreased with the increase of CPUY013 concentration. The novel TOPO Ⅰ inhibitor CPUY013 had a marked inhibition in Capan2 ceils with a distinct dose-dependent manner and induced the apoptosis, probably throught upregulating the expressions of p53, bax, caspase-3 proteins and down regulating the expressions of bcl-2 and TOPO Ⅰ proteins.
出处
《哈尔滨商业大学学报（自然科学版）》
CAS
2008年第2期136-142,共7页
Journal of Harbin University of Commerce：Natural Sciences Edition
