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第三军医大学西南医院全军感染病研究所
纸质出版日期:2008,
网络出版日期:2008-12-30,
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晏泽辉, 邓国宏, 谭文婷, 等. 人肝细胞中雌激素受体α基因启动子使用情况的鉴定[J]. 陆军军医大学学报, 2008,30(24):2276-2280.
YAN ZE-HUI, DENG GUO-HONG, TAN WEN-TING, et al. Identifying the usage of ESR1 promoters in liver cells. [J]. Journal of army medical university, 2008, 30(24): 2276-2280.
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 (PolⅡ-ChIP) to precipitate the promoter sequences and validated them by PCR. Results After PolⅡ-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.
雌激素受体α(ESR1) 启动子RNA多聚酶Ⅱ染色质免疫沉淀
estrogen receptor α promoterRNA Polymerase Ⅱchromatin immunoprecipitation
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