胡椒醛缩氨基均三唑席夫碱对人肝癌细胞分化和凋亡的影响  

Effect of Piperonal Amino-s-Triazole Schiff Base on Differentiation and Apoptosis of Human Hepatocarcinoma Cells

作　　者：房娜[1,2] 马永超[2] 皇甫超申[2] 康玉华[3] 刘彬[2] 屈伸[1] 

机构地区：[1]毕中科技大学同济医学院基础医学院生物化学与分子生物学系,武汉市430030 [2]河南大学医学院、河南大学神经生物学研究所,河南省开封市475004 [3]河南大学淮河临床学院消化科,河南省开封市475001

出　　处：《医学分子生物学杂志》2009年第5期410-414,共5页Journal of Medical Molecular Biology

基　　金：河南省医学科技攻关计划（No.200703080）

摘　　要：目的观察胡椒醛缩氨基均三唑席夫碱对人肝癌细胞BEL-7402的诱导分化和凋亡作用。方法用不同浓度的1－（3，4-二氧亚甲基）苯甲亚氨基－2－吡啶－3－基－5－甲硫基－[1，2，4]三唑（LH－36）与BEL－7402细胞体外培养。MTT法检测细胞增殖和LH－36对细胞增殖的抑制作用。用Hoechst33258荧光染色观察细胞凋亡变化。免疫细胞化学检测细胞内激活型Caspase-3表达。反转录聚合酶链反应（RT—PCR）方法检测细胞甲胎蛋白（α—FP）和白蛋白（Alb）mRNA表达，以了解细胞分化状态。蛋白质免疫印迹分析P53的表达。可见光法测定超氧化物歧化酶（SOD）和过氧化氢酶（CAT）活力。结果LH－36在0．1μmol／L－10μmol／L的浓度范围能抑制细胞增殖，且抑制率随浓度和时间的增加而增高。0．1μmol／L和1．0μmol／L的LH－36使细胞α—FPmRNA表达量明显下降，Alb表达量增高，P53的蛋白表达明显增加，SOD活性升高（P〈0．01），而CAT活性降低，差异有显著性（P〈0．01，P〈0．05）。药物浓度在10μmol／L作用48h，BEL－7402细胞皱缩，呈现凋亡各期改变，Caspase-3阳性细胞明显增加，细胞凋亡率显著高于对照组（P〈0．05）。结论胡椒醛缩氨基均三唑席夫碱对人肝癌BEL-7402细胞具有抗增殖、促分化和凋亡作用，作用可能与过氧化氢的氧化损伤作用有关。Objective To examine the effect of 1- （3, 4-dioxomethylene benzylideneamino） -2- pyridin-3-yl-5-methylsulfanyl- [ 1, 2, 4] -triazole （LH-36） on differentiation and apop- tosis of hepatocarcinoma BEL-7402 cells in vitro. Methods With different concentrations of LH-36 at different time points, BEL-7402 cells were treated in vitro. The effect of LH-36 on cell proliferation and inhibition was confirmed by MTT assay. Apoptotic morphology was determined by Hoechst 33258 labeling. Expression of active caspase-3 in BEL-7402 cells was observed by immunohistochemical staining. Reverse transcriptase polymerase chain reaction （RT-PCR） was used to detect the changes of alpha fetoprotein （α-FP） and albumin （Alb） mRNA in order to explore cellular differentiation state, p53 expression was detected by Western blotting analysis. Colorimetrie method was used to assay the activity of superoxide dismutase （SOD） and catalase （CAT） . Results Cell proliferation was inhibited by LH-36 at 0. 1μmol L^-1 and 10μmol L^-1 in dose- and time-dependent manners. Addition of LH-36 （0. 1μmol L^-1 or 1.0μmol L^-1） increased mRNA expression of Alb and protein expression of p53, but decreased mRNA expression of α-FP. LH-36 markedly promoted the activity of SOD, but not CAT, in BEL-7402 cells. Treatment of BEL-7402 cells with 10μmol L^-1 LH-36 for 48 hours increased the percentage of active caspase-3 positive cells from （3.62 2.87） % to （27.40 2.93 ） % （P 〈 0. 01 ）, and more apoptosis features of cells were observed. Conclusion The anisaldchyde amino-s-triazole schiff base can inhibit proliferation and induce differentiation and apoptosis of human hepatocarcinoma BEL-7402 cells, which might be related to the cytotoxicity of intracellular hydrogen peroxide （H2O2） .

关 键 词：胡椒醛缩氨基均三唑席夫碱 肝癌细胞 细胞分化 细胞凋亡 

分 类 号：R734.2[医药卫生—肿瘤]