当前位置: 首页 > 期刊 > 《新疆医科大学学报》 > 2006年第1期
编号:10936129
低剂量三氧化二砷对HepG2细胞p53基因转录水平的下调作用
http://www.100md.com 《新疆医科大学学报》 2006年第1期
三氧化二砷,,三氧化二砷;p53基因启动子;反式激活;下调,1材料和方法,2结果,3讨论,参考文献:
     (新疆医科大学公共卫生学院, 新疆乌鲁木齐830054; 北京地坛医院, 北京100011; 解放军第302医院传染病研究所基因治疗研究中心, 北京100039)

    摘要: 目的:探讨低剂量三氧化二砷(As2O3)对p53启动子转录活性的调节作用。方法:确定p53启动子区域,聚合酶链反应(PCR)扩增p53p,克隆至真核报告载体pCAT3Basic中,构建pCAT3p53p报告载体;以该质粒转染肝癌细胞系HepG2细胞系,用酶联免疫吸附法(ELISA)检测氯霉素乙酰转移酶(CAT)的表达活性;并以不同浓度三氧化二砷刺激HepG2细胞系,用ELISA法检测CAT的表达活性。结果: 成功克隆出614 bp的新的p53启动子, pCAT3p53p和不同浓度的三氧化二砷(0.25 μmoL/L)瞬时转染的HepG2细胞的CAT表达活性是pCAT3Basic空载体的11.2倍,表明pCAT3p53p具有强的启动子活性。选用0.25、0.5、2.5、5.0 μmol/L浓度的三氧化二砷刺激pCAT3 p53p转染的HepG2细胞,其CAT吸光度值分别为:0.076、0.186、0.149、0.119, CAT活性随着三氧化二砷浓度的倍增呈衰减趋势。结论: p53启动子有反式激活下游基因表达的活性,低剂量三氧化二砷对p53基因转录水平具有下调作用。

    关键词: 三氧化二砷;p53基因启动子;反式激活;下调

    Downregulation of transcriptional activation of p53 gene promoter by low dosage arsenic trioxide

    WU Shunhua, ZHENG Yujian, CHENG Jun, et al

    (Department of Labour and Environment Hygiene, College of Public Health, Xinjiang Medical University, Urumqi 830054, China)

    Abstract: Objective: To investigate the transcriptional activation of p53 gene promoter by low dosage arsenic trioxide (As2O3). Methods: Polymerase chain reaction (PCR) technique was employed to amplify the coding sequence of p53 promoter from HepG2 genomic DNA, and the product was subcloned into pCAT3Basic by Kpn I and Xho I, named pCAT3p53p. pCAT3p53p was transfected into the hepatoblastoma cell line HepG2, then cotransfected with As2O3 by FuGENE 6 transfection reagents. The HepG2 cells transfected with pCAT3Basic was used as negative control. The activity of promoter region fused to the CAT gene in HepG2 cells transfected was detected by a ELISA kit after 48 hours, which reflected the transregulating function of As2O3 to p53p gene promoter. Results: The report vector pCAT3 p53p had been constructed and had been confirmed by restriction enzyme digestion and sequencing. The expression of CAT in HepG2 cells cotransfected with pCAT3 p53p and As2O3 was respectively 0.076 ......

您现在查看是摘要页,全文长 15360 字符