华蟾素注射液对人肝癌HepG2细胞DNA拓扑异构酶Ⅰ的影响.pdfVIP

中国癌症杂志 2010年第20卷第3期 《 》 CHINA ONCOLOGY 2010 Vol.20 No.3 197 华蟾素注射液对人肝癌HepG-2细胞 DNA拓扑异构酶Ⅰ的影响 陈华　孙宇　崔晓楠 大连医科大学第一附属医院肿瘤科，辽宁 大连 116011 　　［摘要］　背景与目的：华蟾素注射液是传统抗肿瘤中药制剂，目前药效机制尚不明确。本研究旨在探讨 华蟾素注射液对DNA拓扑异构酶Ⅰ（topoisomerase  Ⅰ，TOPOⅠ）的影响。方法：采用噻唑蓝还原法（MTT）观 察华蟾素注射液对人肝癌HepG-2细胞增殖的影响；采用流式细胞术（FCM）检测对HepG-2细胞周期的影响；采 用RT-PCR法检测对HepG-2细胞TOPOⅠmRNA表达的影响；采用TOPOⅠ介导的负超螺旋PBR322 DNA解旋反应检测华 蟾素对TOPOⅠ酶活性的影响；采用负超螺旋PBR322 DNA检测华蟾素对DNA的直接抑制作用。结果：华蟾素注射 液对HepG-2细胞增殖具有抑制作用，其抑制效应具有时间和剂量依赖性；并将HepG-2细胞阻滞于S期；RT-PCR 检测表明，华蟾素注射液可下调TOPOⅠmRNA表达，表现浓度依赖效应；对TOPOⅠ介导的负超螺旋PBR322 DNA解 旋反应有抑制作用；对负超螺旋PBR322 DNA没有直接抑制作用。结论：华蟾素注射液能够抑制HepG-2细胞增 殖，影响DNA拓扑异构酶Ⅰ酶的mRNA表达及活性可能为机制之一。 　　［关键词］　华蟾素；　TOPOⅠ；　肝癌；　HepG-2细胞；　PBR322 DNA 　　中图分类号：R73-36+1；R735.7　　文献标识码：A　　文章编号：1007-3639(2010)03-0197-05 The effect of cinobufacini injection on DNA topoisomerase of human hepatocellular carcinoma Ⅰ HepG-2 cells  CHEN Hua, SUN Yu, CUI Xiao-nan (Department of Oncology, The First  Affiliated Hospital of Dalian Medical University, Dalian Liaoning 116011，China) Correspondence to：CUI Xiao-nan　E-mail：cxn23@ 　　［Abstract ］ Background and purpose ：The cinobufacini injection is a traditional antitumor drug. However, its mechanism is still unclear. The purpose of this study was to observe the effect of cinobufacini injections in DNA TOPO of human hepatocellular carcinoma HepG-2 cells. Methods The cells that were proliferated were assessed Ⅰ   ： by MTT assay. Cell cycles were shown through FCM. TOPO mRNA expression was analyzed through RT-PCR. The Ⅰ activity of TOPO was measured by TOPO mediated super coiled PBR322 relaxation. Supercoile