摘要
目的研究芹菜素(API)对人卵巢癌细胞凋亡的影响及其机制。方法培养人卵巢癌SKOV3细胞株,加入不同浓度的API,孵育48 h。PI染色流式细胞术(FCM)分析测定凋亡率;罗丹明染色FCM分析测定细胞线粒体跨膜电位(Δψm);caspase-9分光光度法检测试剂盒测定caspase-9活性;Western印迹检测线粒体凋亡信号转导通路相关蛋白的表达,包括bax、bcl-2、caspase-9和caspase-3。结果API(20,40和80μmol/L)作用48 h能呈浓度依赖性地诱导SKOV3细胞凋亡。而且,API也能降低SKOV3细胞的Δψm,增加caspase-9活性,促进细胞色素c(Cyt c)释放,上调bax、caspase-9和caspase-3蛋白的表达,同时下调bcl-2蛋白表达,且呈剂量依赖性。结论API通过活化线粒体信号转导途径诱导人卵巢癌细胞凋亡。
Objective It is to investigate the influence of apigenin（API） on induction of apoptotic death in human ovarian cancer cells and its underlying mechanisms.Methods Human ovarian cancer SKOV3 cells line was cultured and treated with different concentrations of API for 48 h.The cell apoptosis rat and mitochondrial membrane potential was detected by flow cytometry（FCM） labeled with propidiumiodide（PI） and Rhodamine123 respectively;the caspase-9 activity was determined by caspase-9 colorimetric assay kit;the western blot was used to analyze the expressions of apoptosis mitochondrial signal transduction pathway related proteins including bax,bcl-2,caspase-9 and caspase-3.Results It could concentration-dependently induce the apoptotic death of SKOV3 cell by treatment with API（20,40 and 80μmol/L） for 48 h.Moreover,API could also decrease the cellular Δψm,increase the activity of caspase-9 and enhance the releasing of cytochrome c.The protein expressions of bax,caspase-9 and caspase-3 were upregulated by treatment with API,associated with a downregulation of the protein expression of bcl-2.Conclusion API can induce the apoptosis of human ovarian cancer cells via activating mitochondrial signal transduction pathway.
出处
《现代中西医结合杂志》
CAS
2010年第15期1821-1823,1826,共4页
Modern Journal of Integrated Traditional Chinese and Western Medicine
基金
河北省石家庄市科技攻关课题(08146673)
