三氧化二砷抑制人胆囊癌细胞生长及对细胞周期蛋白D1表达的影响  

Arsenic trioxide inhibits the growth of human gall bladder carcinoma cell and its effect on the cyclin D1

作　　者：凌跃新[1] 艾志龙[1] 锁涛[1] 沈振斌[1] 童赛雄[1] 

机构地区：[1]复旦大学附属中山医院普通外科,上海200032

出　　处：《中国癌症杂志》2007年第11期867-870,共4页China Oncology

基　　金：复旦大学青年科学基金资助(项目编号2006135129)

摘　　要：背景与目的:三氧化二砷(As2O3)在治疗急性早幼粒细胞白血病中取得了突破性成果,近年来它在实体瘤的研究中颇受关注。胆囊癌由于缺乏特异性的临床表现,确诊时多属晚期,手术切除率较低,且常规化疗药物对胆囊癌的疗效较差,因此需寻找治疗胆囊癌新的有效药物。本研究旨在探讨As2O3对人胆囊癌GBC细胞系生长的生物学效应及作用机制。方法:不同浓度As2O3作用于胆囊癌GBC细胞,MTT法检测细胞生长情况,流式细胞仪检测细胞周期分布。Western blot及RT-PCR检测细胞周期蛋白(cyclin)D1、D2、D3、CDK4和CDK6的表达。构建cyclinD1启动子pGL3重组质粒,并转染GBC细胞,测定不同浓度As2O3作用下pGL3相关的荧光素酶活性。结果:As2O3能抑制体外GBC细胞生长,作用呈时间、剂量-效应关系;流式细胞仪检测发现As2O3可将增殖过程中的GBC细胞阻滞于G1期;Western blot及RT-PCR检测显示cyclin D1表达下降;As2O3能降低cyclin D1启动子的表达,作用呈剂量-效应关系,4μmol/L的As2O3处理后cyclin D1的启动子活性下调近70%。结论:As2O3能明显抑制体外人胆囊癌GBC细胞的生长并引起G1期阻滞,主要通过下调cyclin D1的表达来实现。Background and purpose：Arsenic trioxide,verified as a breakthrough in the management of acute promyelocytic leukemia,has been applied to a variety of solid tumors.Gall bladder carcinoma,lacking specific clinical manifestations,is usually diagnosed at advanced stages of the diseases and few cases can be resected by operation.Chemotherapy has not shown significant activity in gall bladder carcinoma.This study was to investigate the biological effect of As2O3 on the growth of human gall bladder carcinoma cell and its mechanism.Methods：GBC cells were cultured with different concentrations of As2O3,the proliferative activity of the cells was detected by MTT methods,and the cell cycle status was carried out by flow cytometry（FCM）.Western blot and RT-PCR were performed to analyse the expression of cyclin D1,D2,D3,CDK4 and CDK6.GBC cells were transient transfected with cyclin D1 promoter construct pGL3 and then treated by different doses of As2O3.The luciferase activity was measured.Results：The treatment of As2O3 in gall bladder carcinoma cells could inhibit the growth of cells in a time and dose dependent manner,make cells arrest in G1 phase and down regulate the expression of cyclin D1.In addition,the activity of cyclin D1 promoter was down-regulated by As2O3 in a dose-dependent manner and decreased about 70 percent when treated with 4 μmol/L As2O3.Conclusions：As2O3 can significantly inhibit the growth of human gall bladder carcinoma cells as well as down-regulate the expression of cyclin D1 in vitro.

关 键 词：三氧化二砷 细胞生长 细胞周期蛋白D1 培养的肿瘤细胞 

分 类 号：R73-36[医药卫生—肿瘤]