国家自然科学基金(30572030) 和湖南省重点学科建设专项基金资助项目.
This work was supported by grants from The National Natural Science Foundation of China (30572030) and The Construct Program of The Key Discipline in Hunan Province.
LCRG1基因( laryngeal carcinoma related gene1,LCRG1 )是一个新的喉癌候选抑瘤基因,其转录调控机制一直未被阐明.通过限制性内切酶酶切介导对LCRG1基因 (-169~+127)区域进行剪切体分析,将LCRG1基因最小启动子定位于-169~-57.应用连接体扫描突变体分析,将关键顺式作用元件确定在-137~-122.生物信息学提示该区存在SP1、E2F1/DP1、EKLF和ZF9转录因子结合位点.利用已知反式作用因子与报告基因质粒进行共转染,提示Spl为有效的反式作用因子,且能上调LCRG1基因的表达.凝胶迁移阻滞实验确定LCRG1基因关键的顺式作用元件区域具有Spl结合位点.LCRG1基因启动子-137~-122片段在该基因表达过程中可能起重要作用,为LCRG1基因功能研究提供了新的证据.
LCRG1(laryngeal carcinoma related gene1, LCRG1), a new candidate tumor suppressor gene of laryngeal carcinoma. However, it is known little about the possible regulatory mechanisms of LCRG1 gene expression. Restriction endonuclease digestion was used to obtain a set of the 5′, or 3′deletion mutants from the region(-169~+127) of the LCRG1 gene. It has been found that the minimal promoter of the LCRG1 gene is mapped at the region from -169~-57. Linker scanning mutational analysis in the region(-169~+127) of the LCRG1 gene was used to identify the crucial cis-elements within the promoter region, The key cis-elements are within the region from -137~-122. SP1, E2F1/DP1, EKLF and ZF9 transcription factor binding site sites were predicted in the region by bioinformatics analysis. Co-transfection with each of a panel of the expression plasmids of the known transcription factors with the relevant reporter construct indicates Sp1 is potent transcription factor for enhancement of the promoter activity, SP1 can also up-regulate the endogenous expression of LCRG1 gene. Electrophoretic mobility shift assay (EMSA) was applied to verify that the key cis-elements of LCRG1 gene exist sequence of Sp1 binding sites. The findings, which showed that the key cis-elements within the region from -137~-122 play an important role in expression of the LCRG1 gene, provide a novel evidence for further study of the function of LCRG1 gene.
谢海龙,陈主初,李金花,曾龙武,谭桂煌.LCRG1基因启动子关键顺式调节元件的鉴定[J].生物化学与生物物理进展,2009,36(5):633-640
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