文章摘要
杨璐西 翟东霞 张丹英 俞超芹 蔡在龙.Mir-26a 调控子宫肌瘤中孕激素受体a(PRa)、雌激素受体α(ER-alpha) 的表达[J].,2016,16(21):4019-4023
Mir-26a 调控子宫肌瘤中孕激素受体a(PRa)、雌激素受体α(ER-alpha) 的表达
Mir-26a Regulate ER-α, PRa Expression in Leiomyoma Uteri
  
DOI:
中文关键词: ERα  PRa  miR-26a  子宫肌瘤
英文关键词: ER-α  PRa  mir-26a  leiomyoma uteri
基金项目:国家自然科学基金项目重点项目(30930113)
作者单位
杨璐西 翟东霞 张丹英 俞超芹 蔡在龙 兰州大学第二医院 第二军医大学附属长海医院中心实验室第二军医大学附属长海医院中医科 
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中文摘要:
      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)、雌激素受体琢(ER琢)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-26a mimics 于子宫肌瘤平滑肌细胞,western blotting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ER琢、 PRa 为mir-26a 的靶基因。蛋白水平进一步验证,mir-26a mimics 的转染量不同,ER琢、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 抑制雌激素受体琢(ER琢)、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琢(ER琢)、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。
英文摘要:
      Objective:Estrogen and progesterone play an important role in pathogenesis of uterine fibroids. The role of miRNA in the pathogenesis of uterine fibroids is still poorly understood. The preliminary experiment verify that mir-26a is lower expressed in uterine fibroids. This study aimed to further study regulatory effect of mir-26a on ERα, PRa of leiomyoma uteri in vitro.Methods:The potential target genes of mir-26a were predicted by Targetscan. The fragment of ER琢, PRa 3'UTR which contained the binding site of mir-26a 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-26a 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-26a. After verification, mir-26a mimics were transfected into uterine fibroids muscle cells. The expression levels of ERα, PRa were detected by western blotting.Results:TargetScan and dual luciferase reporter gene detection system further confirmed that mir-26a regulated ERα, PRa mR- NA in the level of post transcription (P<0.05). ERα, PRa protein level were down regulated when mir-26a was transfected into leiomyo- ma uteri cells(P<0.05). Different transfection amount of mir-26a mimics resulted in the different level of ERalpha and PRa protein expres- sion.Conclusion:Mir-26a binding site was at the 3'UTR of target genes. The expression of ERα, PRa may be inhibited by mir-26a in leiomyoma uteri. Mir-26a may affect the development of leiomyoma uteri by regulating ERαand PRa. This study verified the mechanics of mir-26a in leiomyoma uteri. It is the foundmental of leiomyoma uteri treatment, On the other hand decreases the operative trauma.
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