土木香乙酸乙酯提取物对人胰腺癌Capan-2细胞增殖的抑制作用及机制研究  

Study on the Inhibitory Effect and Mechanism of Inula helenium Ethyl Acetate Extract on Proliferation of Human Pancreatic Cancer Capan-2 Cells

作　　者：王霖玲 曾健梅 阎优优 张博 林能明 

机构地区：[1]浙江中医药大学附属杭州第一医院转化医学研究中心,杭州310006 [2]杭州市第一人民医院/南京医科大学附属杭州医院转化医学研究中心,杭州310006

出　　处：《中国药房》2017年第31期4384-4388,共5页China Pharmacy

基　　金：浙江省卫生高层次创新人才培养工程项目(No.2010-190-4)

摘　　要：目的:研究土木香乙酸乙酯提取物(IHE)对人胰腺癌Capan-2细胞增殖的抑制作用及机制。方法:采用MTT法测定0、0.5、1、2、4、8μg/mL的IHE作用48 h后细胞的增殖抑制率,克隆形成试验观察0、1、2μg/mL的IHE作用1周后对细胞克隆形成的影响,Hoechst 33342染色法观察0、2、4μg/mL的IHE作用48 h后细胞核形态的变化,流式细胞术检测0、4、8、16μg/mL的IHE作用48 h后细胞的凋亡率,JC-1染色法观察0、4、8、16μg/mL的IHE作用24 h后细胞线粒体膜电位变化,Western blot法检测0、4、8、16μg/mL的IHE作用48 h后细胞线粒体凋亡相关蛋白Bcl-2、Bax、Mcl-1和p53上调凋亡因子(PUMA)、多聚二磷酸腺苷核糖聚合酶(PARP)蛋白的表达。结果:2、4、8μg/mL的IHE对细胞的增殖有明显抑制作用,且呈浓度依赖性,半数抑制浓度为6.6μg/mL;1、2μg/mL的IHE可明显抑制细胞的克隆形成;4μg/mL的IHE可明显造成细胞核固缩;8、16μg/mL的IHE可明显促进细胞凋亡,16μg/mL的IHE作用48 h后细胞凋亡率达到45.53%;16μg/mL的IHE作用24 h后可引起82.47%的细胞线粒体膜电位下降;8μg/mL的IHE可明显下调细胞中Bcl-2、Mcl-1、PUMA、PARP蛋白表达,16μg/mL的IHE可明显下调细胞中Mcl-1、PUMA表达。结论:IHE可能通过引起细胞线粒体膜电位的下降,下调细胞中PUMA、Mcl-1蛋白的表达,引起细胞的凋亡,从而发挥其抑制人胰腺癌Capan-2细胞增殖的作用。OBJECTIVE：To study the inhibitory effect and mechanism of Inula helenium ethyl acetate extract（IHE）on proliferation of human pancreatic cancer Capan-2 cells. METHODS：MTT was used to determine the cell proliferation inhibition rate after treated by 0,0.5,1,2,4,8 μg/mL IHE;clone formation test was used to observe the effects of 0,1,2 μg/mL IHE treating for 1 week on cell clone formation;Hoechest 33342 staining was used to observe the changes of nuclear morphology after treated by 0,2,4 μg/mL IHE for 48 h;flow cytometry was used to detect the cell apoptosis rate after treated by 0,4,8,16 μg/mL IHE for 48 h;JC-1 staining was used to observe the changes of intracellular mitochondrial membrane potential after treated by 0,4,8,16 μg/mL IHE for 24 h;Western blot was used to detect the expressions of mitochondrial apoptosis-related proteins Bcl-2,Bax,Mcl-1,p53 up-regulated modulator of apoptosis（PUMA）,and polymerase（PARP）after treated by 0,4,8,16 μg/mL IHE for48 h. RESULTS：2,4,8 μg/mL IHE had obvious inhibitory effect on cell proliferation,showing concentration-dependent relationship,with IC（50） of 6.6 μg/mL;1,2 μg/mL IHE can obviously inhibit the clone formation of cells;4 μg/mL IHE can obviously cause cell nuclear condensation;8,16 μg/mL IHE can obviously promote the cell apoptosis,and the cell apoptosis rate reached 45.53% after treated by 16 μg/mL IHE for 48 h;16 μg/mL IHE treating for 24 h can cause the decrease of 82.47% cells＇ mitochondrial membrane potential;8 μg/mL IHE can obviously down-regulate the protein expressions of Bcl-2,Mcl-1,PUMA and PARP,and 16 μg/mL IHE can obviously down-regulate the expressions of Mcl-1 and PUMA. CONCLUSIONS：IHE may show itsinhibitory effect on proliferation of human pancreatic cancer Capan-2 cells by causing the decrease of mitochondrial membrane potential in cells and down-regulating the protein expressions of Mcl-1 and PUMA to cause cell apoptosis.

关 键 词：土木香 乙酸乙酯提取物 人胰腺癌Capan-2细胞 线粒体 细胞凋亡 

分 类 号：R285.5[医药卫生—中药学]