The regulation of the transcription factor STAT1 to 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；通过染色质免疫沉淀（chromatin immunoprecipitation，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及蛋白表达的影响。结果： Gas6基因在乳腺癌中高表达；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，并且有调控Gas6启动子活性的功能性结合位点。转录因子STAT1可在转录水平对癌症相关基因Gas6进行正向调控。结论：癌症相关基因Gas6是转录因子STAT1的直接靶基因。
Abstract:
Objective：The current study aims to investigate the effect and mechanism of signal transducer and activator of transcription 1（STAT1）on cancer-related gene Gas6. Methods：The database was used 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 chromatin immunoprecipitation（ChIP）assays. The binding sites of STAT1 on Gas6 promoter were predicted by bioinformatics analysis and identified by mutation plasmid construction，and the effects of binding site mutation on activity of Gas6 promoter was detected by 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 blot，respectively. Results：Gas6 was a highly expressed gene in breast cancer. Gas6 was positively associated with STAT1 in multiple cancers. The Gas6 promoter contained binding sites of STAT1. STAT1 positively regulateed Gas6 at transcriptional levels. Conclusion：The cancer related gene Gas6 is a direct target gene for transcription factor STAT1.
