孕激素通过调节 H19 和 miR-152 的表达来调节 Wnt 和 β-catenin 信号通路，从而改善子宫内膜息肉。

Progesterone ameliorates the endometrial polyp by modulating the signaling pathway of Wnt and β-catenin via regulating the expression of H19 and miR-152.

机构信息

Reproductive Medicine Department, Xianyang Central Hospital, Xianyang, Shaanxi, China.

Department of Gynecology and Obstetrics, Baoji Maternal and Child Health Hospital, Baoji, Shaanxi, China.

出版信息

J Cell Biochem. 2019 Jun;120(6):10164-10174. doi: 10.1002/jcb.28301. Epub 2019 Jan 18.

DOI:10.1002/jcb.28301

PMID:30659641

Abstract

BACKGROUND

MicroRNAs and long noncoding RNAs are believed to play important roles in the pathogenesis of various diseases. This study aimed to explore the potential mechanism of the involvement of H19 and miR-152 in an endometrial polyp.

METHODS

Luciferase assay was conducted to determine the effect of progesterone. Real-time polymerase chain reaction (PCR) and western blot were performed to detect the influence of progesterone on miR-152 and Wnt1. MTT assay and flow cytometry (FCM) were utilized to detect the effect of progesterone on cell proliferation and apoptosis. In silicon analysis, luciferase assay, real-time PCR, and immunohistochemistry (IHC) were performed to explore the regulatory relationship between H19 and miR-152 or miR-152 and Wnt1.

RESULTS

Progesterone dose-dependently increased the H19 expression level through driving the promoter efficiency of H19. Then, progesterone upregulated Wnt1 level and downregulated miR-152 in a dose-dependent manner in ECC1 and HEC1A cells. Administration of progesterone inhibited cell viability and promoted cell apoptosis. H19 negatively regulated miR-152 expression by binding to miR-152. Furthermore, Wnt1 was identified as a virtual target gene of miR-152 and was inhibited by miR-152. Progesterone receptors mRNA and miR-152 were lowly expressed in participants with an endometrial polyp, while the levels of H19 and Wnt1 were much higher in the endometrial polyp group compared with normal controls. H19 negatively regulated miR-152 and miR-152 negatively regulated Wnt1, with the negative correlation coefficients being -0.500 and -0.500, respectively. Using IHC, it was found that Wnt1 and Bcl-2 protein were highly expressed in the endometrial polyp group compared with normal controls.

CONCLUSION

The results suggested that H19 was associated with endometrial polyp via mediating cell proliferation and apoptosis.

摘要

背景

MicroRNAs 和长链非编码 RNA 被认为在各种疾病的发病机制中发挥重要作用。本研究旨在探讨 H19 和 miR-152 参与子宫内膜息肉的潜在机制。

方法

通过荧光素酶报告基因实验确定孕激素的作用。实时聚合酶链反应（PCR）和 Western blot 用于检测孕激素对 miR-152 和 Wnt1 的影响。MTT 检测和流式细胞术（FCM）用于检测孕激素对细胞增殖和凋亡的影响。通过荧光素酶报告基因实验、实时 PCR 和免疫组化（IHC）探讨 H19 与 miR-152 或 miR-152 与 Wnt1 之间的调控关系。

结果

孕激素通过驱动 H19 启动子效率，呈剂量依赖性增加 H19 的表达水平。然后，孕激素在 ECC1 和 HEC1A 细胞中呈剂量依赖性地上调 Wnt1 水平并下调 miR-152 水平。孕激素给药抑制细胞活力并促进细胞凋亡。H19 通过与 miR-152 结合负调控 miR-152 的表达。此外，Wnt1 被鉴定为 miR-152 的虚拟靶基因，并被 miR-152 抑制。孕激素受体 mRNA 和 miR-152 在子宫内膜息肉患者中低表达，而 H19 和 Wnt1 的水平在子宫内膜息肉组中明显高于正常对照组。H19 负调控 miR-152，而 miR-152 负调控 Wnt1，负相关系数分别为-0.500 和-0.500。通过 IHC 发现，Wnt1 和 Bcl-2 蛋白在子宫内膜息肉组中的表达明显高于正常对照组。