雌激素对载脂蛋白AⅠ基因启动子不同区域转录活性的影响  

The Effect of Estradiol on Modulating Transcription of Different Fractions of ApoA Ⅰ Gene Promoter

作　　者：崔丽[1,2] 毛用敏[3] 赵莉莉[3] 程爱娟[3] 崔让庄[3] 

机构地区：[1]天津医科大学,300070 [2]天津市南开医院 [3]天津市胸科医院

出　　处：《天津医药》2011年第10期896-898,共3页Tianjin Medical Journal

摘　　要：目的:探讨雌激素对载脂蛋白(Apo)AⅠ基因启动子不同区域转录调节活性的影响。方法:利用含荧光素酶报告基因的表达载体pGL2构建携带ApoAⅠ基因启动子不同区域片段的重组质粒和同时携带ApoCⅢ/AⅣ基因片段的重组质粒。阳离子脂质体法将重组质粒与pRL-null内参质粒共转染HepG2细胞,加入10μmol/L雌激素刺激24h检测细胞荧光素酶报告基因的荧光强度,以反映ApoAⅠ的转录水平。结果:pGL2/-256AⅠ,pGL2/-2500AⅠ,pGL2/-256AⅠCⅢAⅣ及pGL2/-2500AⅠCⅢAⅣ转染后雌激素组荧光素酶相对活性明显高于对照组(P<0.05);而pGL2/-41AⅠ及pGL2/-41AⅠCⅢAⅣ质粒转染后雌激素组与对照组荧光素酶活性差异无统计学意义(P>0.05)。结论:ApoAⅠ启动子-256^-41区域可能包含与雌激素作用相关的反应元件,受雌激素刺激后转录活性增强。Objective： To analyze the modulating mechanism of estrogen on different fractions of apolipoprotein （apo） A Ⅰ gene promoter. Methods： The recombinants carrying different length DNA fragments of Apo A Ⅰ gene promoter with or without a 7 kb region ApoC Ⅲ/AⅣ intergenic region in basic pGL2 vector that containing lueiferase reporter gene were constructed. The final recombinants were transfected with pRL-null vector as an internal control to HepG2 ceils by the cationic lipid method. The cells were treated with estradiol at 10μmol/L concentration for 24 h, and then cells were collected and analysed. The activities of these luciferase enzymes were measured with Dual-GloTM Luciferase Assay System. Results： The recombinant expressions of lueiferase enzymes including pGL2/-256AⅠ, pGL2-2500AⅠ , pGLd-256A Ⅰ C Ⅲ A Ⅳ and pGLd-2500A ⅠC ⅢAⅣ were significantly increased in estradiol induced experiments, exception of the recombinant plasraids containing only the basal promoter （-41/＋397 bp） with or without region of Apo C Ⅲ/A Ⅳ compared with those of control group（P 〈 0.05）. Conclusion： The region of-256 bp to-41 bp in Apo AⅠ promoter may be the smallest reactive element for estrogen, which can increase transcription activity after estrogen stimulation.

关 键 词：载脂蛋白A-Ⅰ 启动区(遗传学)调节元件 转录雌二醇质粒转染细胞系 肿瘤 

分 类 号：R285[医药卫生—中药学]