Mir-26a调控子宫肌瘤中孕激素受体a(PRa)、雌激素受体α(ERα)的表达  

Mir-26a Regulate ERα, PRa Expression in Leiomyoma Uteri

作　　者：杨璐西[1,2] 翟东霞[3] 张丹英[3] 俞超芹[3] 蔡在龙[2] 

机构地区：[1]兰州大学第二医院,甘肃兰州730000 [2]第二军医大学附属长海医院中心实验室,上海200433 [3]第二军医大学附属长海医院中医科,上海200433

出　　处：《现代生物医学进展》2016年第21期4019-4023,共5页Progress in Modern Biomedicine

基　　金：国家自然科学基金项目重点项目(30930113)

摘　　要：目的:雌激素和孕激素在子宫肌瘤发病中起重要作用。但miRNA在子宫肌瘤发病中的作用还知之甚少,我们前期已证实mir-26a在子宫肌瘤中低表达,本实验进一步探讨mir-26a在体外对子宫肌瘤中孕激素受体a(PRa)、雌激素受体α(ERα)表达的调控。方法:利用TargetScan软件预测mir-26a的潜在靶基因,找出靶基因3'UTR区片段,插入PmirGLO绿色荧光蛋白编码区下游,构建报告基因载体,同时原代培养子宫肌瘤平滑肌细胞。将报告基因载体与mir-26a共转染入原代培养的子宫肌瘤平滑肌细胞,引入双荧光素酶报告基因系统对mir-26a的靶基因进行验证。转染mir-26amimics于子宫肌瘤平滑肌细胞,westernblotting检测子宫肌瘤平滑肌细胞中mir-26a靶蛋白表达水平。结果:用TargetScan软件和双荧光素酶报告基因系统证实ERα、PRa为mir-26a的靶基因。蛋白水平进一步验证,mir-26amimics的转染量不同,ERα、PRa的蛋白表达水平下调不同。结论:Mir-26a通过结合靶基因的3'-UTR区调控靶基因的mRNA水平。Mir-26a抑制雌激素受体α(ERα)、孕激素受体a(PRa)在子宫肌瘤中的表达。Mir-26a可能通过调控雌激素受体α(ERα)、孕激素受体a(PRa)影响子宫肌瘤的发展。本实验通过确定mir-26a对子宫肌瘤的作用机制,有望进一步提高子宫肌瘤的治疗技术,减少手术治疗的创伤。Objective： Estrogen and progesterone play an important role in pathogenesis of uterine fibroids. The role of mi RNA in the pathogenesis of uterine fibroids is still poorly understood. The preliminary experiment verify that mir-26 a is lower expressed in uterine fibroids. This study aimed to further study regulatory effect of mir-26 a on ERα, PRa of leiomyoma uteri in vitro. Methods： The potential target genes of mir-26 a were predicted by Targetscan. The fragment of ERα, PRa 3＇UTR which contained the binding site of mir-26 a was inserted into the plasmid named Pmir GLO, respectively. A reporter gene vector was constructed. Leiomyoma uteri cells were primary cultured in vitro. The reporter gene vectors and mir-26 a were co-transfected into primary cultured smooth muscle cells of uterine fibroids. Dual-Luciferase reporter gene detection system was used to confirm that ERα, PRa were the target genes of mir-26 a.After verification, mir-26 a mimics were transfected into uterine fibroids muscle cells. The expression levels of ERα, PRa were detected by western blotting. Results： Target Scan and dual luciferase reporter gene detection system further confirmed that mir-26 a regulated ERα, PRa m R- NA in the level of post transcription（P〈0.05）. ERα, PRa protein level were down regulated when mir-26 a was transfected into leiomyo- ma uteri cells（P〈0.05）. Different transfection amount of mir-26 a mimics resulted in the different level of ERα and PRa protein expres- sion. Conclusions： Mir-26 a binding site was at the 3＇UTR of target genes. The expression of ERα, PRa may be inhibited by mir-26 a in leiomyoma uteri. Mir-26 a may affect the development of leiomyoma uteri by regulating ERα and PRa. This study verified the mechanics of mir-26 a in leiomyoma uteri. It is the foundmental of leiomyoma uteri treatment, On the other hand decreases the operative trauma.

关 键 词：ERΑ PRA miR-26a 子宫肌瘤 

分 类 号：R-33[医药卫生] Q78[生物学—分子生物学]