雌激素上调LRP16基因表达的研究

Study on expression upregulation of LRP16 induced by estrogen

  • 摘要: 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 (serial analysis of gene expression)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,luciferase assay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。

     

    Abstract: Objective: To explore and identify the regulatory way of LRP16 gene expression. Methods: Cis-elements of LRP16gene promoter region and SAGE(Serial Analysis of Gene Expression)pattern of LRP16were analysed,the results suggested that 17β-estradiol(β-E2) may upregulate its expression.To prove the hypothesis,the human LRP16gene promoter, with 2.6kb of 5'-flanking DNA, was cloned upstream of the Luciferase gene (pS0) and tested for estrogen regulation by transient cotransfection with estrogen receptor α or β(ERα,ERβ)gene expression vector in COS-7 and MCF-7cells.The relative Luciferase activity was detected by Luciferase Assay.Results: The relative luciferase activity was significantly increased in MCF-7 and COS-7cells cotransfected with pS0 and ERβexpression vector than those with pS0transfection and pS0 and ERβ expression vector cotransfection, and the increased extent in MCF-7 and COS-7cells 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-7is valuable to be further investigated.

     

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