急性髓系白血病中Nras基因点突变的检测以及信号传导通路蛋白的表达.doc

急性髓系白血病中Nras基因点突变的检测以及信号传导通路蛋白的表达 　　作者：杨建刚,孙雪梅,顾香芳,徐祖琼　　作者单位：1.东南大学 临床医学院，江苏 南京 210009;2.南京大学医学院附属南京鼓楼医院血液科，江苏 南京 210008 　　【摘要】目的 研究Nras基因在急性髓系白血病(AML)中的突变以及在AML和正常对照组中RAS蛋白下游信号传导通路上AKT、ERK1/2以及其活化形式PAKT和PERK1/2的表达情况。方法 应用聚合酶链反应单链构象多态性法(PCRSSCP)及直接测序法检测AML中Nras基因突变率，应用Western blot法检测AKT、ERK以及其活化形式PAKT,PERK1/2的表达量。结果 48例AML中检测到2例Nras 12突变，2例Nras 61突变，突变组PAKT、PERK1/2的表达水平较正常对照组明显升高(P0.05)。未有Nras基因突变的AML其PAKT、PERK1/2的表达水平也增高，但表达不均一。结论 Nras基因突变可能是AML发生发展的原因之一，其机制可能是由于突变导致编码的Ras蛋白构型改变，使其处于功能持续活化状态，从而激活了Raf/Mek/Erk和PI3K/Akt等下游信号传导通路，最终导致AML的发生。 　　【关键词】 Nras基因,急性髓系白血病,突变;PAKT;PERK 　　Abstract：Objective To study the mutation rate of Nras gene in acute myelocytic leukemia (AML) and the effect of Nras gene on the RAS protein downriver signal conduction circuit AKT,ERK1/2 and its activation form PAKT and the PERK1/2 expression in AML and the normal group.Methods Polymerase chain reactionsingle strand conformation polymorphism(PCRSSCP) and direct sequencing of AML were utilized to detect Nras gene mutation rate.Western blot was employed to detect AKT,ERK and PAKT,PERK1/2 expression levels.Results In 48 patients with AML,two cases of Nras 12 mutation were detected,as well as two cases of Nras 61 mutation.In the mutation group,PAKT,PERK1/2 expression levels increased significantly than normal control group(P0.05).In the cases of AML with no Nras gene mutation,the expressions of PAKT,PERK1/2 also increased,but not to the uniform extent.Conclusion Nras gene mutation may play a role in the development of AML;It causes the mutations in the Ras protein coding gene,leading to structure changes and constitutive activation of Ras protein.Activated Ras protein then activates Raf/Mek/Erk and PI3K/Akt downstream signal transduction pathways,eventually leading to the incidence of AML. 　　Keywords:Nras gene;acute myelocytic leukemia;mutation;PAKT;PERK 　　越来越多的研究表明，急性髓系白血病(AML)的发病与基因突变有关，其突变过程由多步骤完成，目前已经得到公认的是二次打击学说。这些多步骤基因改变往往最终归结到共同的信号转导和转录途径，如Ras信