对甲氧基肉桂醛左氧氟沙星酰腙诱导人肝癌SMMC-7721细胞凋亡的作用  

p-Methoxycinnamaldehyde levofloxacin-3-ylhydrazone induces apoptosis of human hepatocarcinoma SMMC-7721 cells

作　　者：付建民[1,2] 王德辉[2] 孙金平[1] 康玉华[1] 皇甫超申[2] 胡国强[3] 刘彬[1] 

机构地区：[1]河南大学护理学院神经生物学研究所,河南开封475004 [2]河南大学医学院,河南开封475004 [3]河南大学化学生物学研究所,河南开封475004

出　　处：《中国药理学通报》2011年第6期801-805,共5页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No20872028);河南省教育厅自然科学研究计划项目(No2009B310001;2010B310002);开封市科技发展计划项目(No080312);河南大学科技攻关项目(No07YBGG004)

摘　　要：目的探讨喹诺酮酰腙类化合物诱导人肝癌SMMC-7721细胞凋亡的作用。方法用不同浓度的对甲氧基肉桂醛左氧氟酰腙(FQ-10)与SMMC-7721细胞体外培养。MTT法检测FQ-10对SMMC-7721细胞的增殖抑制作用;Hoechst33258/PI双染荧光染色法、TUNEL法及琼脂糖凝胶电泳检测细胞凋亡变化;高内涵活细胞成像系统测定细胞线粒体膜电位(△ψm)变化;Western blot方法测定caspase-9、caspase-8、caspase-3、p53、Bcl-2、Bax蛋白表达。结果 FQ-10在0.625～10.00μmol.L-1的浓度范围能抑制细胞增殖,呈时间、浓度依赖性;作用于细胞24 h的IC50值是4.40μmol.L-1;各组FQ-10作用24 h后,细胞凋亡率高于对照组(P<0.05);琼脂糖凝胶电泳可见凋亡细胞典型的梯状DNA条带,并伴有线粒体膜电位降低。荧光染色观察细胞形态学变化,显示凋亡细胞染色质凝集,细胞核碎裂成碎片等典型细胞凋亡特征性变化,晚期凋亡细胞可见特异性PI染色。与对照组比较,FQ-10作用后p53、Bax、caspase-9、caspase-3蛋白表达量增加,其中caspase-9、caspase-3活性裂解片段明显增加,caspase-8无变化,而Bcl-2蛋白表达水平下降。结论对甲氧基肉桂醛左氧氟酰腙能够激活线粒体凋亡通路,诱导人肝癌细胞凋亡。Aim To study the effect of（S） N-（3-p-methoxylphenyl）-6-fluoro-7-（4-methylpiperazin-1-yl）-1,8-（3,1-propoxy）-quinolin-4（1H）-one-3-carbonyl hydrazine（FQ-10） on apoptosis of hepatocarcinoma SMMC-7721 cells in vitro.Methods With different concentrations of FQ-10 at different times used to treat SMMC-7721 cells in vitro,the proliferation of the cells and the inhibition effect of FQ-10 on the cell proliferation were examined by MTT assay.Cell apoptosis was determined by Hoechst 33258/PI fluorescence staining,TUNEL and agarose gel electrophoresis method.Mitochondrial membrane potential（MMP,△ψm） was measured by high content screening image system.The caspase-9,caspase-8,caspase-3,p53,Bcl-2 and Bax protein expressions were detected by Western blot analysis.Results The cell proliferation was inhibited by FQ-10 at 0.625~10.00 μmol·L-1 in a time-and-dose dependent manner.Treatment of SMMC-7721 cells with different concentrations of FQ-Henan 10 for 24 h increased the percentage of the apoptosis cells（P0.05）.The typical ladder DNA in apoptotic cells and a concomitant dissipation of the mitochondrial membrane potential could be observed by agarose gel electrophoresis.Typical cell apoptotic features could be seen and specific PI staining was observed in late apoptotic cells.In addition,FQ-10（3.0~7.39 μmol·L-1） increased protein expression of p53,Bax,caspase-9,caspase-3,separately,and induced cytosolic accumulation of activities caspase-9 and caspase-3,whereas protein expression of Bcl-2 decreased with no change of caspase-8.Conclusion The mitochondrial-dependent pathways were involved in FQ-10 induction of apoptosis of SMMC-7721 cells.

关 键 词：氟喹诺酮酰腙 肝癌细胞 细胞增殖 凋亡 线粒体膜电位 p53 

分 类 号：R329.24[医药卫生—人体解剖和组织胚胎学] R329.25[医药卫生—基础医学]