华蟾素注射液对人肝癌HepG-2细胞增殖及拓扑异构酶表达的影响  

Effects of Cinobufacini Injections on Proliferation and Expression of Topoisomerase of Hepatocarcinoma HepG-2 Cells

作　　者：高山[1] 田莉莉[1] 陈华[1] 崔晓楠[1] 

机构地区：[1]大连医科大学第一附属医院肿瘤科,辽宁大连116011

出　　处：《中国实验方剂学杂志》2013年第11期250-255,共6页Chinese Journal of Experimental Traditional Medical Formulae

基　　金：国家自然科学基金项目(81173615)

摘　　要：目的:探讨华蟾素注射液对人肝癌HepG-2细胞增殖及拓扑异构酶Ⅰ(TopoⅠ)和Ⅱ的影响。方法:实验分为对照组(不加药)与实验组(华蟾素注射液0.105,0.21,0.42 mg.L-1);采用四甲基偶氮唑蓝还原法(MTT)观察对HepG-2细胞增殖的影响;采用流式细胞术(AnnexinV/PI)双染色法检测对HepG-2细胞凋亡的影响;采用流式细胞术法检测对HepG-2细胞周期的影响;采用逆转录-多聚酶联反应(RT-PCR)法检测对肝癌HepG-2细胞TopoⅠ,TopoⅡmRNA水平表达的影响;采用蛋白免疫印迹法(Western-blot法)观察对HepG-2细胞中TopoⅠ,TopoⅡ蛋白表达的影响。结果:华蟾素注射液抑制HepG-2细胞增殖(P<0.05),诱导细胞凋亡(P<0.05),呈时间和剂量依赖性;阻滞HepG-2细胞于S期(P<0.05),呈剂量依赖性;下调TopoⅠ,TopoⅡmRNA(P<0.05)及蛋白的表达(P<0.05),呈剂量依赖性。结论:华蟾素注射液能够抑制人肝癌HepG-2细胞增殖,诱导癌细胞凋亡,下调TopoⅠ,TopoⅡ表达可能是其机制之一。Objective： To study the effects of cinobufacini injections on the proliferation and expression of topoisomerase of hepatocarcinoma HepG-2 cells. Method： Experiment were divided into control group and experimental group （cinobufacini injections group 0.105,0.21,0.42 mg·L^-1）.Cell proliferation was assessed by Methyl Thiazolyl Tetrazolium blue reduction （MTT）assay.Apoptosis was detected by Annexin V/PI staining. Cell cycles were shown through flow cytometry method （FCM）. TopoisomeraseⅠ（TopoⅠ）,TopoisomeraseⅡ （Topo Ⅱ） mRNA were analyzed by Reverse transcription-polymerase chain reaction （RT-PCR）. Expressions of TopoⅠ,Topo Ⅱ protein were analyzed by Western-blot. Result： Cinobufacini injection significantly inhibited HepG-2 cellproliferation（P〈0.05）,induced apoptosis（P〈0.05） in a dose and time dependent manner,induced tumor cells arrested at the S-phase a dose dependent manner.Down regulated mRNA and protein expressions of TopoⅠ,Topo Ⅱ（P〈0.05）in a dose dependent manner. Conclusion： Cinobufacini injections can inhibit human hepatocellular carcinoma HepG-2 cells proliferation,induced apoptosis and induced tumor cells arrested at the S-phase.The down regulation of TopoⅠ,Topo Ⅱ expressions might be one of the anti-tumor mechanisms of cinobufacini injection.

关 键 词：华蟾素注射液 HEPG-2细胞 增殖 凋亡 拓扑异构酶Ⅰ 拓扑异构酶Ⅱ 

分 类 号：R285.5[医药卫生—中药学]