转录因子YY1对口腔鳞癌细胞ITGB6基因表达的影响

发布时间：2018-04-04 01:39

本文选题：YY转录因子　切入点：整合素β　出处：《中国病理生理杂志》2014年03期

【摘要】：目的:研究肿瘤相关转录因子YY1对口腔鳞癌细胞整合素β6(integrinβ6,ITGB6)基因表达调控的影响。方法:用生物信息学方法预测分布在ITGB6启动子区域的转录因子YY1潜在的结合位点,构建萤光素酶报告基因质粒,利用双萤光素酶报告基因系统检测ITGB6启动子片段的转录活性;利用染色质免疫沉淀技术检测在天然染色质条件下转录因子YY1与ITGB6启动子的结合情况;采用定点突变方法检测YY1结合位点对ITGB6启动子活性的影响;利用RT-PCR方法检测过表达转录因子YY1对口腔鳞癌细胞ITGB6 mRNA表达水平的影响。结果:ITGB6启动子-421~-150 nt区域存在多个转录因子YY1潜在的结合位点。在口腔鳞癌细胞的天然染色质中,转录因子YY1结合于ITGB6启动子-421~-150 nt区域;定点突变YY1潜在结合位点对口腔鳞癌细胞中ITGB6启动子活性无显著影响。另外,过表达的YY1对口腔鳞癌细胞ITGB6 mRNA的表达水平也无影响。结论:转录因子YY1在口腔鳞癌细胞中结合于ITGB6基因启动子-421~-150 nt区域,但对ITGB6基因的基础转录水平无影响。
[Abstract]:Aim: to investigate the effect of tumor related transcription factor (YY1) on integrin 尾 6(integrin 尾 6 ITGB6 gene expression in oral squamous carcinoma cells.Methods: the potential binding sites of transcription factor YY1 in ITGB6 promoter region were predicted by bioinformatics method. The plasmids of luciferase reporter gene were constructed and the transcriptional activity of ITGB6 promoter fragment was detected by double luciferase reporter gene system.Chromatin immunoprecipitation technique was used to detect the binding of transcription factor YY1 to ITGB6 promoter under natural chromatin condition, and the effect of YY1 binding site on ITGB6 promoter activity was detected by site-directed mutation method.The effect of overexpression of transcription factor YY1 on the expression of ITGB6 mRNA in oral squamous carcinoma cells was detected by RT-PCR method.Results there were many potential binding sites of transcription factor YY1 in the region of -421 ~ 150 NT of the 1: ITGB6 promoter.In the natural chromatin of oral squamous carcinoma cells, transcription factor YY1 binds to the region of ITGB6 promoter -421 ~ (-150 NT), while site-directed mutagenesis YY1 potential binding site has no significant effect on the activity of ITGB6 promoter in oral squamous cell carcinoma cells.In addition, overexpression of YY1 had no effect on the expression of ITGB6 mRNA in oral squamous carcinoma cells.Conclusion: transcription factor YY1 binds to the region of ITGB6 gene promoter -421 ~ (-1) -150 NT in oral squamous cell carcinoma cells, but has no effect on the basic transcription level of ITGB6 gene.
【作者单位】：厦门市口腔医院;汕头大学医学院细胞衰老实验室;厦门大学医学院基础医学部;
【基金】：国家自然科学基金资助项目(No.30900661;No.81072208;No.81370160) 教育部留学回国人员科研启动基金资助项目汕头大学医学院基础与临床科研基金资助项目
【分类号】：R739.85

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