转录因子STAT1对癌症相关基因Gas6的调控研究
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南京医科大学第一附属医院儿科

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国家自然科学基金项目(面上项目,重点项目,重大项目)


The regulation of the transcription factor STAT1 to human cancer-related gene Gas6
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    摘要:

    目的:研究信号转导和转录激活因子1(signal transducer and activator of transcription 1,STAT1)对癌症相关生长抑制特异性基因6(growth arrest specific 6,Gas6)的影响及其机制。方法:应用数据库预测乳腺癌的基因聚类分析结果,并预测Gas6与STAT1在多种癌症中的相关性;通过ChIP实验检测STAT1是否可与Gas6启动子结合,应用数据库预测STAT1在Gas6启动子上的结合位点并突变其关键碱基,通过双萤光素酶报告基因实验检测结合位点突变对Gas6启动子活性的影响,同时分别检测敲低或过表达STAT1对Gas6启动子和突变的Gas6启动子活性的影响;通过实时荧光定量PCR技术和蛋白质免疫印迹技术分别检测敲低或过表达STAT1对 Gas6 mRNA及蛋白质表达的影响。结果: Gas6是乳腺癌的高表达基因;Gas6与STAT1在多种癌症中有正相关趋势;Gas6启动子序列包含STAT1的结合位点,并且有可对Gas6启动子活性调控的功能性结合位点。转录因子STAT1可在转录水平对癌症相关基因Gas6正向调控。结论:癌症相关基因Gas6是转录因子STAT1的直接靶基因。

    Abstract:

    Objective: The effect and mechanism of STAT1 on cancer-related gene Gas6 were investigated. Methods: Using the database to predict the results of the gene cluster analysis of breast cancer. And to predict the correlation between Gas6 and STAT1 in multiple cancers. The binding of STAT1 to Gas6 promoter was tested by ChIP assays. The binding sites of STAT1 on Gas6 promoter were predicted by bioinformatics analysis. The binding site was identified by mutation plasmid construction and dual luciferase reporter assay. The effects of knockdown and overexpression of STAT1 on Gas6 mRNA and protein expression were measured by qRT-PCR and Western blotting, respectively. Results: Gas6 is a highly expressed gene in breast cancer. Gas6 is positively associated with STAT1 in multiple cancers. The Gas6 promoter contains binding sites of STAT1. STAT1 positively regulates Gas6 at transcriptional levels. Conclusion: Gas6 is a direct target gene of transcription factor STAT1.

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  • 收稿日期:2023-03-07
  • 最后修改日期:2023-05-20
  • 录用日期:2023-10-25
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