【作者】 孙佳；

【导师】 叶丽红；

【作者基本信息】 南京中医药大学 ， 中医学， 2012， 硕士

【摘要】 研究目的：探讨国医大师经验方“抗癌扶正方”中泽漆的单体槲皮素、没食子酸在抗肝癌、促进细胞凋亡方面的作用机制。研究方法：体外培养肝癌细胞SMMC-7721,顺铂为阳性组,观察不同浓度槲皮素、没食子酸对细胞凋亡的作用及其相关机制。采用MTT法检测槲皮素、没食子酸对细胞生长的影响,并观察在不同时间点的细胞形态；利用流式细胞仪检测药物作用后的细胞凋亡情况及细胞周期分布；运用RT-PCR及Western Blot法检测单体对SMMC-7721细胞PI3K/AKT信号通路相关凋亡蛋白表达的影响。研究结果：槲皮素、没食子酸对人肝癌细胞SMMC-7721增殖有明显抑制作用,呈浓度依赖性和时间依赖性。流式结果提示,槲皮素可将SMMC-7721细胞周期阻滞于G0/G1期,并且其凋亡率随浓度增加而逐渐增加；没食子酸可使SMMC-7721细胞周期阻滞于Go/G1期,凋亡率随药物浓度增加的上升趋势不显著。细胞形态观察,药物组细胞呈现不规则形,甚至变圆,体积缩小,细胞结构没有对照组紧密,某些细胞甚至出现脱落、漂浮现象。RT-PCR、Western Blot司亡相关蛋白表达检测表明,药物组AKT的表达受抑制,PTEN、Caspase9蛋白的表达率随着药物浓度的增加而增加。研究结论：槲皮素、没食子酸均能诱导人肝癌细胞SMMC-7721凋亡,其机制可能是使细胞周期阻滞于G0/G1期,PTEN的过表达抑制AKT活化,激活Caspase9从而促进细胞凋亡。PI3K/AKT言号通路的表达,可能是槲皮素、没食子酸诱导肝癌细胞SMMC-7721凋亡的机制之一。更多还原

【Abstract】 Objective:Discussion Zeqi’s monomer Quercetin, Gallic acid of the ALC of GuoYi master experience prescriptions inhibit liver cancer and promote in cell apoptosis of the role of mechanism.Methods:In vitro liver cancer cells SMMC-7721,cisplatin for positive group,observe different concentration Quercetin,Gallic acid in cell apoptosis role and its related mechanism.By MTT method to detect Quercetin,Gallic acid on the influence of the cell growth,and observed in different time points cells form;Use of flow cytometric detection after cell apoptosis of drugs and cell cycle distribution;Use RT-PCR and Western Blot method to detect monomer to SMMC-7721cells PI3K/AKT signaling pathways related apoptosis the influence of protein expression.Results:Quercetin,Gallic acid to human liver cancer cells SMMC-7721value-added had significant inhibitory effect,show concentration dependence and time dependence.Streaming suggests,Quercetin can make the SMMC-7721in cell cycle block G0/G1phase,and the apoptosis rate increased with the concentration;Gallic acid,can make the SMMC-7721in cell cycle block G0/G1phase,apoptosis rate of drug concentration increases with the increasing trend is not significant.Cell morphology,drug group of cells present irregular form,or even go round,the volume is narrow,the cellular structure no control group closely,some cells appear even fall off,floating phenomenon.RT-PCR,the Western Blot apoptosis related protein expression check showed that the expression of the drug group AKT inhibition,PTEN,Caspase9protein expression rate with the drug concentration increased.Conclusion:Quercetin,Gallic acid,all can induce human liver cell apoptosis SMMC-7721,its mechanism may be to block the cell cycle in G0/G1phase,the PTEN gene expression AKT activation had inhibition,activation, Caspase9and promote cell apoptosis.P13K/AKT signal pathways of expression,may be Qucrcetin,Gallic acid induced liver cancer cells SMMC-7721apoptosis of one of the mechanism.更多还原