Wnt通路抑制因子Dickkopf-1的表达作用  

Effect on expression of Wnt signaling pathway inhibitor Dickkopf-1

作　　者：腊蕾[1] 班武[2] 饶进军[2] 吴曙光[3] 

机构地区：[1]南方医科大学南方医院药学部,广东广州510515 [2]广州军区广州总医院,广东广州510010 [3]南方医科大学药学院,广东广州510515

出　　处：《中国医院药学杂志》2007年第5期629-632,共4页Chinese Journal of Hospital Pharmacy

摘　　要：目的:研究Wnt通路抑制因子Dickkopf-1(DKK-1)的表达作用。方法:将携带p53基因的复制缺陷型腺病毒载体(Adp53)导入到p53完全缺失的人肝癌细胞株Hep3B中,并以Wnt通路的关键因子β-链接蛋白(-βcatenin)的改变为功能指标评价Wnt通路的变化。以逆转录聚合酶链反应(RT-PCR)技术检测Wnt通路抑制因子DKK-1表达的调节作用,以流式细胞术检测Adp53的转基因情况和-βcatenin的表达。结果:DKK-1mRNA水平在转染Adp53 20h后即有明显升高32h达最高水平,随后逐渐降低。量效关系研究表明在转染Adp53剂量为0.5,5,50pfu/cell时DKK1mRNA表达均有显著增高,尤以50pfu/cell时表达水平最高。-βcatenin表达水平随着转染时间和转染剂量的增加,阳性细胞百分比强度和平均荧光量强度表达水平逐渐下降。结论:外源性p53能够诱导Wnt通路抑制剂DKK-1的表达,进而产生抑制Wnt通路的作用。OBJECTIVE To investigate the effect of Wnt signaling pathway inhibitor DKK-1. METIHODS Human hepatocarcinoma Hep3B ceils were transfected with a replication-defective advenovirus encoding（Adp53）, and Wnt signaling pathway function changes with key factor β-catenin. DKK-1 mRNA expression detected by reverse transfection（ RT）-PCR, β-catenin and Adp53 transgene expression was measured by fluorescence-activated cell sorting（FACS）. RESULTS DKK-1 mRNA in Hep3B ceils was initially potentiated by transfection with Adp53 after 20 h, and the peak expression was seen at 32 h. Dose-effect studies revealed that Adp53 in dose range of 0. 5,5 and 50 pfu/cetl could promote the expression of DKK-1 mRNA,with the highest expression at the dose of 50 pfu/celL ,8-catenin, positive cells percent and fluorescence intensity was descent , along with increased transfection time and dosage gain. CONCLUSION Ectogentic p53 promotes mRNA expression of Wnt signaling pathway inhibitor DKK-1 in Hep3B and suppress Wnt signaling pathway.

关 键 词：Adp53 DICKKOPF-1 WNT通路 β-链接蛋白 

分 类 号：R394.6[医药卫生—医学遗传学] Q756[医药卫生—基础医学]