论文部分内容阅读
探讨乙酰水杨酸增强多胺衍生物ANISpm抗肝癌作用及其作用机制。采用MTT法检测细胞毒性;应用高内涵活细胞成像系统检测细胞内ANISpm含量的变化及对细胞凋亡和线粒体膜电位的影响;采用Western blotting方法检测细胞色素c,精脒/精胺乙酰转移酶,Caspase-3、-8、-9等蛋白表达的变化;应用高效液相色谱法检测细胞内多胺含量的变化。应用H22肿瘤移植模型评价对实体瘤生长的影响。实验结果显示,乙酰水杨酸体内外均具有增强ANISpm抗肝癌的作用,其机制为乙酰水杨酸通过上调肝癌细胞内精脒/精胺乙酰转移酶的活性,降低细胞内多胺含量从而增加了ANISpm的摄取,并通过ANISpm诱导肝癌细胞凋亡。该凋亡作用与ANISpm诱导细胞线粒体膜电位降低,促进细胞色素c释放以及活化Caspase-3、-8、-9等有关。
To investigate the anti-hepatocarcinoma effect of acetylsalicylic acid on polyamine derivative ANISpm and its mechanism. MTT assay was used to detect the cytotoxicity. The content of ANISpm and the effect on cell apoptosis and mitochondrial membrane potential were detected by high-content living cell imaging system. The levels of cytochrome c, spermine / spermine acetyltransferase , Caspase-3, -8, -9 and other protein expression changes; application of high performance liquid chromatography detection of intracellular polyamine content changes. H22 tumor transplantation model evaluation of the impact of solid tumor growth. The experimental results show that acetylsalicylic acid has the anti-hepatocarcinoma effect of ANISpm both in vitro and in vivo. The mechanism is that acetylsalicylic acid increases the activity of spermine / spermine acetyltransferase in hepatoma cells and decreases the content of intracellular polyamines Uptake of ANISpm and induction of hepatoma cell apoptosis by ANISpm. This apoptosis is related to the decrease of mitochondrial membrane potential, the release of cytochrome c and the activation of Caspase-3, -8, -9 in ANISpm.