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
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篇名: 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
TITLE:
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、8 μg/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、2 μg/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、4 μg/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、16 μg/mL的IHE作用48 h后细胞的凋亡率,JC-1染色法观察0、4、8、16 μg/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、16  μg/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、8 μg/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6.6 μg/mL;1、2 μg/mL的IHE可明显抑制细胞的克隆形成;4 μg/mL的IHE可明显造成细胞核固缩;8、16 μg/mL的IHE可明显促进细胞凋亡,16    μg/mL的IHE作用48 h后细胞凋亡率达到45.53%;16 μg/mL的IHE作用24 h后可引起82.47%的细胞线粒体膜电位下降;8 μg/mL的IHE可明显下调细胞中Bcl-2、Mcl-1、PUMA、PARP蛋白表达,16 μg/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。
ABSTRACT: 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 for 48 h. RESULTS: 2, 4, 8 μg/mL IHE had obvious inhibitory effect on cell proliferation, showing concentration-dependent relationship, with IC50 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 its inhibitory 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.
期刊: 2017年第28卷第31期
作者: 王霖玲,曾健梅,阎优优,张博,林能明
AUTHORS: WANG Linling,ZENG Jianmei,YAN Youyou,ZHANG Bo,LIN Nengming
关键字: 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
KEYWORDS: Inula helenium; Ethyl acetate extract; Human pancreatic cancer Capan-2 cells; Mitochondrion; Cell apoptosis
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