【作者】 刘冉； 尹立红； 浦跃朴； 张文翠； 胡旭； 刘耀珍；

【Author】 LIU Ran, YIN Li-hong, PU Yue-pu, ZHANG Wen-cui, HU Xu, LIU Yao-zhen (School of Public Health, Southeast University, Nanjing 210009,China; Chuzhou Center for Disease Prevention and Control, Huaian 223200, China)

【机构】 东南大学公共卫生学院； 江苏省淮安市楚州区疾病预防控制中心；

【摘要】 [目的]建立检测CYP2E1 mRNA的SYBR Green I实时定量PCR的方法,了解正常人外周血中CYP2E1的表达水平。[方法]采集25例随机个体的外周血样本,提取PBMC中的RNA;制备CYP2E1定量模板,采刚SYBR GREEN 1实时荧光定量逆转录-聚合酶链反应(RT-PCR)检测CYP2E1基因的表达。[结果]标准曲线循环阈值与模板浓度具有良好的线性关系,R2为0.998。25例随机个体的CYP2E1 mRNA的表达范围在1.26×104-5.30×105拷贝/μg总RNA。[结论]应用SYBR Green I实时定量检测CYP2E1基因表达水平,为研究CYP2E1表达调控的机制、表达水平和肿瘤的关系具有重要意义。更多还原

【Abstract】 [Objective] To establish a SYBR Green I quantitative real-time PCR method for detecting the expression of CYP2E1 gene in Human peripheral blood monocytes(PBMC). [Methods] RNA was isolated from PBMC of 25 Human blood sample. CYP2E1 PCR product was purified for the standard template. The level of the CYP2E1 gene expression was measured using real time RT-PCR with SYBR Green I. [Results] The standard curve indicated the linear relationship between CT(cycle threshold) template concentration and showed high correlations (R2= 0.998) . CYP2E1 expression levels varied from 1.26×104 to 5.30×105 mRNA copies/μg total RNA. [Conclusions] CYP2E1 mRNA was successfully detected by SYBR Green I quantitative real-time PCR. It is important for this method to be used for further study of CYP2E1 mRNA expression regulation mechanism and the relationship with carcinoma.更多还原