论文部分内容阅读
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 promoter of estrogen receptor α (ESR1) gene in human hepatocytes. Methods HepG2 and L02 cells were cultured and stimulated routinely, cells were fixed with formalin and sonicated. Chromatin immunoprecipitation (PolⅡ-ChIP) was carried out by using Pol Ⅱ CTD-specific Ser5 specific antibody. Eight potential promoter regions Primers were designed for PCR amplification to identify the use of ESR1 gene promoter. Results Pol II-ChIP was found after β-estradiol stimulated different hepatocytes. Pol II-binding fragments of ESR1 promoter E1, E2 and F were precipitated in HepG2 hepatoma cell line, but only promoter D Pol II bound fragments precipitated. Conclusion The expression of ESR1 in different hepatocytes is regulated by different promoters. The expression of ESR1 in HepG2 hepatocarcinoma cell lines is regulated by promoters E1, E2 and F, while the expression of ESR1 in L02 cell line is regulated by promoter D.