雌激素上调LRP16基因表达的研究

Study on expression upregulation of LRP16 induced by estrogen

目的 :鉴定LRP16基因的表达调控途径并探讨其可能机制。方法 :对LRP16基因进行启动子序列顺式作用元件和信息学角度的SAGE(serialanalysisofgeneexpression)谱分析 ,提示雌激素对其可能具有转录调控作用。为证实这一作用 ,构建了LRP16基因启动子序列 (2 6kb)调控的荧光素酶报告子 (pS0 ) ,并与雌激素受体α和β(ERα ,ERβ)真核表达载体共转染COS 7和MCF 7细胞 ,加入雌二醇 (β E2 )培养 ,luciferaseassay方法测定相对荧光素酶活性。结果 :报告子 pS0 与ERα真核表达载体共转染细胞的相对荧光素酶活性较非共转染组及 pS0 /ERβ表达载体共转染组显著升高 ,并且在两种细胞中的升高幅度接近。结论 :LRP16是受雌激素调控的一个新识别的靶基因 ,具体调控途径由雌激素变构激活的ERα直接介导 ,其临床意义有待进一步研究。

Objective:To explore and identify the regulatory way of LRP16 gene expression Methods: Cis elements of LRP16 gene promoter region and SAGE(Serial Analysis of Gene Expression)pattern of LRP16 were analysed, the results suggested that 17β estradiol(β E 2 ) may upregulate its expression To prove the hypothesis, the human LRP16 gene promoter, with 2 6kb of 5' flanking DNA, was cloned upstream of the Luciferase gene (pS 0 )and tested for estrogen regulation by transient cotransfection with estrogen receptor α or β(ERα,ERβ)gene expression vector in COS 7 and MCF 7 cells The relative Luciferase activity was detected by Luciferase Assay Results: The relative luciferase activity was significantly increased in MCF 7 and COS 7 cells cotransfected with pS 0 and ERβ expression vector than those with pS 0 transfection and pS 0 and ERβ expression vector cotransfection, and the increased extent in MCF 7 and COS 7 cells was nearly consistent Conclusion: LRP16 is a novelly identified target gene regulated by estrogen The regulatory approach is mediated by ERα The potential clinical significance of expression upregulation of LRP16 gene induced by estrogen in breast cancer cell line MCF 7 is valuable to be further investigated