人肝细胞中雌激素受体α基因启动子使用情况的鉴定  

Identifying the usage of ESR1 promoters in liver cells

作　　者：晏泽辉[1] 邓国宏[1] 谭文婷[1] 刘国栋[1] 但芸婕[1] 王宇明[1] 

机构地区：[1]第三军医大学西南医院全军感染病研究所,重庆400038

出　　处：《第三军医大学学报》2008年第24期2276-2280,共5页Journal of Third Military Medical University

基　　金：国家自然科学基金(30470964;30671850);国家重点基础研究发展计划973计划(2007CB512900)~~

摘　　要：目的鉴定人肝细胞中雌激素受体α(estrogen receptorα,ESR1)基因启动子使用情况。方法常规培养和刺激HepG2和L02细胞,甲醛固定细胞,超声剪切染色质,采用利用PolⅡ CTD末端Ser5特异性抗体进行染色质免疫沉淀(PolⅡ-ChIP),针对文献报道的8个可能启动子区设计引物进行PCR扩增,鉴定ESR1基因启动子的使用情况。结果β-雌二醇刺激不同肝细胞后进行PolⅡ-ChIP发现,HepG2肝癌细胞株中ESR1启动子E1、E2、F沉淀出有PolⅡ的结合的片段,而在L02细胞株中则仅启动子D有PolⅡ的结合的片段沉淀出来。结论不同肝细胞中ESR1的表达受不同的启动子的调控。HepG2肝癌细胞株中ESR1的表达受启动子E1、E2、F调控,而在L02细胞株中ESR1的表达受启动子D调控。Objective To identify the usage of ESR1 promoters in two kinds of liver cells. Methods We cultured and stimulated HepG2 and L02 cells by routine methods. After the cells were cross-linked by formaldehyde and chromatin was sonicated by ultrasonic, we performed RNA polymerase Ⅱ-chromatin immunoprecipitations （Pol Ⅱ-CHIP） to precipitate the promoter sequences and validated them by PCR. Results After Pol Ⅱ- ChIP assay for the liver cells stimulated by β-estradiol, E1, E2 and F promoters of ESR1 could be precipitated in HepG2 cells, while only D promoter could be precipitated in L02 cells. Conclusion We found that the transcription and expression of ESR1 in different liver cells are regulated by different promoters, like what we found ESR1 expression in HepG2 cells by E1, E2 and F promoters, ESR1 expression in L02 cell by D promoter.

关 键 词：雌激素受体α(ESR1) 启动子 RNA多聚酶Ⅱ 染色质免疫沉淀 

分 类 号：R394-33[医药卫生—医学遗传学] R394.3[医药卫生—基础医学]