Graham Amanda, Falcone Tommaso, Nothnick Warren B
Department of Molecular and Integrative Physiology, University of Kansas Medical Center, Kansas City, KS 66160, USA Institute for Reproductive Health and Regenerative Medicine, Center for Reproductive Sciences, University of Kansas Medical Center, Kansas City, KS 66160, USA.
Department of Department of Obstetrics, Gynecology and Women's Health Institute, Cleveland Clinic, Cleveland, OH 44195, USA.
Hum Reprod. 2015 Mar;30(3):642-52. doi: 10.1093/humrep/dev005. Epub 2015 Jan 29.
What is the role of microRNA-451 (miR-451) in human endometriotic tissue?
miR451 expression was elevated in endometriotic lesion tissue. MiR451 modulated the expression of macrophage migration inhibitory factor and limited cell survival.
microRNAs are post-transcriptional regulators of gene expression which have been reported to be mis-expressed in endometriotic tissue. The exact pattern of expression and role of miR451 in endometriosis is currently unknown.
STUDY DESIGN, SIZE, DURATION: Thirty women with endometriosis are included in the study.
PARTICIPANTS/MATERIALS, SETTING, METHODS: Matched eutopic (N = 30) and endometriotic lesion tissue (N = 43) were collected. miR-451, macrophage migration inhibitory factor (MIF), cyclin E1 (CCNE) and phosphatase and tensin homolog (PTEN) mRNA expression were examined by quantitative real-time (qRT)-PCR while MIF protein expression was evaluated by western blot analysis. miR-451 regulation of MIF in vitro translation was confirmed by 3'untranslated region (UTR) reporter assays and western blot analysis. The effect of miR-451 on cell survival was assessed using a human endometrial epithelial cell line (HES).
Compared with eutopic endometrium, both MIF mRNA and protein were significantly (P < 0.05) decreased in endometriotic lesions and this was associated with a significant (P < 0.05) increase in miR-451 expression. Transfection of HES cells with luciferase reporter constructs for MIF revealed that miR-451 specifically bound to the 3'UTR to regulate expression. Further, forced expression of miR-451 induced a significant (P < 0.05) down-regulation of both MIF mRNA and protein in HES cells which was associated with a significant (P < 0.05) reduction in cell survival. Inhibition of MIF using a specific antagonist verified that reduction of MIF contributes to HES cell survival.
LIMITATIONS, REASONS FOR CAUTION: miR-451 and MIF expression were only examined in tissue from women with endometriosis.
Our data support the hypothesis that miR-451 is elevated in endometriotic tissue and, through regulating MIF expression, may function to limit endometriotic lesion cell survival.
STUDY FUNDING/COMPETING INTERESTS: This study was funded by the National Institutes of Health/NICHD by grant NIH HD069043 to W.B.N. The authors have no competing interests.
微小RNA-451（miR-451）在人子宫内膜异位组织中起什么作用？
miR451在子宫内膜异位病变组织中的表达升高。miR451调节巨噬细胞迁移抑制因子的表达并限制细胞存活。
微小RNA是基因表达的转录后调节因子，据报道在子宫内膜异位组织中表达异常。miR451在子宫内膜异位症中的具体表达模式和作用目前尚不清楚。
研究设计、规模、持续时间：本研究纳入了30名子宫内膜异位症患者。
研究对象/材料、环境、方法：收集了配对的在位内膜组织（n = 30）和子宫内膜异位病变组织（n = 43）。通过定量实时（qRT）-PCR检测miR-451、巨噬细胞迁移抑制因子（MIF）、细胞周期蛋白E1（CCNE）和磷酸酶及张力蛋白同源物（PTEN）的mRNA表达，同时通过蛋白质印迹分析评估MIF蛋白表达。通过3'非翻译区（UTR）报告基因检测和蛋白质印迹分析证实了miR-451在体外对MIF翻译的调节作用。使用人子宫内膜上皮细胞系（HES）评估miR-451对细胞存活的影响。
与在位内膜相比，子宫内膜异位病变中MIF的mRNA和蛋白水平均显著降低（P < 0.05），这与miR-451表达的显著升高（P < 0.05）相关。用MIF的荧光素酶报告基因构建体转染HES细胞表明，miR-451特异性结合3'UTR以调节表达。此外，miR-451的强制表达导致HES细胞中MIF的mRNA和蛋白水平显著下调（P < 0.05），这与细胞存活率的显著降低（P < 0.05）相关。使用特异性拮抗剂抑制MIF证实MIF的降低有助于HES细胞存活。
局限性、谨慎的原因：仅在子宫内膜异位症患者的组织中检测了miR-451和MIF的表达。
我们的数据支持以下假设，即miR-451在子宫内膜异位组织中升高，并通过调节MIF表达，可能起到限制子宫内膜异位病变细胞存活的作用。
研究资金/利益冲突：本研究由美国国立卫生研究院/国家儿童健康与人类发展研究所资助，授予W.B.N.的拨款NIH HD069043。作者没有利益冲突。
