The Long Noncoding RNA MEG3 Regulates Ras-MAPK Pathway Through RASA1 in Trophoblast and is Associated with Unexplained Recurrent Spontaneous Abortion

Overview

Journal Mol Med

Publisher Biomed Central

Specialties General Medicine
Molecular Biology

Date 2021 Jul 9

PMID 34238211

Citations 14

Authors

Jun Zhang

Xinqiong Liu

YaLi Gao

Affiliations

Soon will be listed here.

Abstract

Background: Maternally Expressed Gene 3 (MEG3) is expressed at low levels in placental villi during preeclampsia; however, its roles in unexplained recurrent spontaneous abortion (URSA) remain unclear. In this study, we aimed to explore the relationship between MEG3 and URSA.

Methods: The differentially expressed lncRNAs (MEG3) and its downstream genes (RASA1) were identified using bioinformatics analysis of Genomic Spatial Event (GSE) database. The expression levels of MEG3 in embryonic villis (with gestational ages of 49-63 days) and primary trophoblasts were determined using quantitative RT-PCR assay. A mouse model of Embryo implantation, Cell Counting Kit-8 (CCK-8), flow cytometry, and Transwell migration assays were performed to determine the implantation, proliferative, apoptotic, and invasive capacities of trophoblast. The level of phosphorylated core proteins in the RAS-MAPK pathway were analyzed using Western blot assay. The mechanisms of MEG3 in the regulation of RASA1 were studied by RNA pulldown, RNA immunoprecipitation (RIP), DNA pulldown, and chromatin immunoprecipitation (ChIP) assays.

Results: MEG3 had a low expression level in embryonic villis of 102 URSA patients compared with those of 102 normal pregnant women. MEG3 could promote proliferation and invasion, inhibit the apoptosis of primary trophoblast of URSA patients (PT-U cells), as well as promote embryo implantation of mouse. Besides, MEG3 also promoted the phosphorylation of rapidly accelerated fibrosarcoma (Raf), mitogen-activated protein kinase kinase (MEK), and extracellular-signal-regulated kinase (ERK) proteins. The results of RNA pull down and RIP assays showed that MEG3 bound with the enhancer of zeste homolog 2 (EZH2). The DNA pulldown assay revealed that MEG3 could bind to the promoter sequence of the RAS P21 Protein Activator 1 (RASA1) gene. Further, the ChIP assay showed that MEG3 promoted the binding of EZH2 to the promoter region of the RASA1 gene.

Conclusions: The inactivation of MEG3 in embryonic villi association with URSA; MEG3 inhibited the expression of RASA1 by mediating the histone methylation of the promoter of RASA1 gene by EZH2, thereby activating the RAS-MAPK pathway and enhancing the proliferative and invasive capacities of trophoblasts.

Citing Articles

Transcriptomic effects of alginate hydrogel applied to the production of bovine embryos.

Ferronato G, Rosa P, Bridi A, Dos Santos A, Nociti R, Chiaratti M Heliyon. 2025; 10(24):e40957.

PMID: 39759294 PMC: 11700250. DOI: 10.1016/j.heliyon.2024.e40957.

Circular RNAs as a novel class of potential therapeutic and diagnostic biomarkers in reproductive biology/diseases.

Song W, Chen X, Wu H, Rahimian N Eur J Med Res. 2024; 29(1):643.

PMID: 39741306 PMC: 11689626. DOI: 10.1186/s40001-024-02230-7.

Silencing of maternally expressed RNAs in Dlk1-Dio3 domain causes fatal vascular injury in the fetal liver.

Yu H, Zhao Y, Cheng R, Wang M, Hu X, Zhang X Cell Mol Life Sci. 2024; 81(1):429.

PMID: 39382697 PMC: 11465015. DOI: 10.1007/s00018-024-05462-2.

Maternal RNA transcription in Dlk1-Dio3 domain is critical for proper development of the mouse placental vasculature.

Zhang X, He H, Yu H, Teng X, Wang Z, Li C Commun Biol. 2024; 7(1):363.

PMID: 38521877 PMC: 10960817. DOI: 10.1038/s42003-024-06038-3.

Deciphering the Epigenetic Landscape: Placental Development and Its Role in Pregnancy Outcomes.

Chen Y, Ye Z, Lin M, Zhu L, Xu L, Wang X Stem Cell Rev Rep. 2024; 20(4):996-1014.

PMID: 38457061 DOI: 10.1007/s12015-024-10699-2.

References

Gao Y, Lu X . Decreased expression of MEG3 contributes to retinoblastoma progression and affects retinoblastoma cell growth by regulating the activity of Wnt/β-catenin pathway. Tumour Biol. 2015; 37(2):1461-9. DOI: 10.1007/s13277-015-4564-y. View

Zeng X, Hunt A, Jin S, Duran D, Gaillard J, Kahle K . EphrinB2-EphB4-RASA1 Signaling in Human Cerebrovascular Development and Disease. Trends Mol Med. 2019; 25(4):265-286. PMC: 6456402. DOI: 10.1016/j.molmed.2019.01.009. View

Guo J, Yin L, Chen Y, Jin X, Zhou X, Zhu N . Autologous blood transfusion augments impaired wound healing in diabetic mice by enhancing lncRNA H19 expression via the HIF-1α signaling pathway. Cell Commun Signal. 2018; 16(1):84. PMC: 6245761. DOI: 10.1186/s12964-018-0290-6. View

Gao Y, Huang P, Zhang J . Hypermethylation of MEG3 promoter correlates with inactivation of MEG3 and poor prognosis in patients with retinoblastoma. J Transl Med. 2017; 15(1):268. PMC: 5747938. DOI: 10.1186/s12967-017-1372-8. View

Meng W, Cui W, Zhao L, Chi W, Cao H, Wang B . Aberrant methylation and downregulation of ZNF667-AS1 and ZNF667 promote the malignant progression of laryngeal squamous cell carcinoma. J Biomed Sci. 2019; 26(1):13. PMC: 6347788. DOI: 10.1186/s12929-019-0506-0. View

Zhang J, Yao T, Wang Y, Yu J, Liu Y, Lin Z . Long noncoding RNA MEG3 is downregulated in cervical cancer and affects cell proliferation and apoptosis by regulating miR-21. Cancer Biol Ther. 2015; 17(1):104-13. PMC: 4847830. DOI: 10.1080/15384047.2015.1108496. View

Zhang J, Yao T, Lin Z, Gao Y . Aberrant Methylation of MEG3 Functions as a Potential Plasma-Based Biomarker for Cervical Cancer. Sci Rep. 2017; 7(1):6271. PMC: 5524906. DOI: 10.1038/s41598-017-06502-7. View

Tang L, Liang Y, Xie H, Yang X, Zheng G . Long non-coding RNAs in cutaneous biology and proliferative skin diseases: Advances and perspectives. Cell Prolif. 2019; 53(1):e12698. PMC: 6985680. DOI: 10.1111/cpr.12698. View

Chu C, Qu K, Zhong F, Artandi S, Chang H . Genomic maps of long noncoding RNA occupancy reveal principles of RNA-chromatin interactions. Mol Cell. 2011; 44(4):667-78. PMC: 3249421. DOI: 10.1016/j.molcel.2011.08.027. View

10.

Wang L, Tang H, Xiong Y, Tang L . Differential expression profile of long noncoding RNAs in human chorionic villi of early recurrent miscarriage. Clin Chim Acta. 2016; 464:17-23. DOI: 10.1016/j.cca.2016.11.001. View

11.

Wang Y, Xie Y, Li L, He Y, Zheng D, Yu P . EZH2 RIP-seq Identifies Tissue-specific Long Non-coding RNAs. Curr Gene Ther. 2018; 18(5):275-285. PMC: 6249712. DOI: 10.2174/1566523218666181008125010. View

12.

Kaneko S, Bonasio R, Saldana-Meyer R, Yoshida T, Son J, Nishino K . Interactions between JARID2 and noncoding RNAs regulate PRC2 recruitment to chromatin. Mol Cell. 2013; 53(2):290-300. PMC: 4026005. DOI: 10.1016/j.molcel.2013.11.012. View

13.

Zhang J, Gao Y . Long non-coding RNA MEG3 inhibits cervical cancer cell growth by promoting degradation of P-STAT3 protein via ubiquitination. Cancer Cell Int. 2019; 19:175. PMC: 6615085. DOI: 10.1186/s12935-019-0893-z. View

14.

Zhang Y, Zou Y, Wang W, Zuo Q, Jiang Z, Sun M . Down-regulated long non-coding RNA MEG3 and its effect on promoting apoptosis and suppressing migration of trophoblast cells. J Cell Biochem. 2014; 116(4):542-50. DOI: 10.1002/jcb.25004. View

15.

Zhang Y, Luo G, Zhang Y, Zhang M, Zhou J, Gao W . Critical effects of long non-coding RNA on fibrosis diseases. Exp Mol Med. 2018; 50(1):e428. PMC: 5799794. DOI: 10.1038/emm.2017.223. View

16.

Yu X, Hu L, Li S, Shen J, Wang D, Xu R . Long non-coding RNA Taurine upregulated gene 1 promotes osteosarcoma cell metastasis by mediating HIF-1α via miR-143-5p. Cell Death Dis. 2019; 10(4):280. PMC: 6433912. DOI: 10.1038/s41419-019-1509-1. View

17.

Zhou H, Tang K, Liu H, Zeng J, Li H, Yan L . Regulatory Network of Two Tumor-Suppressive Noncoding RNAs Interferes with the Growth and Metastasis of Renal Cell Carcinoma. Mol Ther Nucleic Acids. 2019; 16:554-565. PMC: 6506628. DOI: 10.1016/j.omtn.2019.04.005. View

18.

Li C, Tan F, Pei Q, Zhou Z, Zhou Y, Zhang L . Non-coding RNA MFI2-AS1 promotes colorectal cancer cell proliferation, migration and invasion through miR-574-5p/MYCBP axis. Cell Prolif. 2019; 52(4):e12632. PMC: 6668983. DOI: 10.1111/cpr.12632. View

19.

Chanda K, Das S, Chakraborty J, Bucha S, Maitra A, Chatterjee R . Altered Levels of Long NcRNAs Meg3 and Neat1 in Cell And Animal Models Of Huntington's Disease. RNA Biol. 2018; 15(10):1348-1363. PMC: 6284602. DOI: 10.1080/15476286.2018.1534524. View

20.

Horita K, Kurosaki H, Nakatake M, Kuwano N, Oishi T, Itamochi H . lncRNA UCA1-Mediated Cdc42 Signaling Promotes Oncolytic Vaccinia Virus Cell-to-Cell Spread in Ovarian Cancer. Mol Ther Oncolytics. 2019; 13:35-48. PMC: 6463205. DOI: 10.1016/j.omto.2019.03.003. View