miR‑424‑5p is downregulated in the placentas of patients with preeclampsia and affects trophoblast migration and invasion

Zhu,Weimin; Chen,Xing

doi:10.3892/etm.2023.11993

miR‑424‑5p is downregulated in the placentas of patients with preeclampsia and affects trophoblast migration and invasion

Authors:
- Weimin Zhu
- Xing Chen
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Affiliations: Department of Gynecology and Obstetrics, The Obstetrics and Gynecology Hospital of Fudan University, Shanghai 200011, P.R. China
Published online on: May 5, 2023 https://doi.org/10.3892/etm.2023.11993
Article Number: 294
Copyright: © Zhu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Insufficient invasion of trophoblast cells has been reported to be closely associated with the pathogenesis of preeclampsia (PE). MicroRNAs (miRs) have essential roles in the trophoblasts invasion via targeting specific genes with diverse functions. However, the underlying mechanism remains largely unclear and requires further investigation. The present study aimed to identify and evaluate the potential functions of miRs in trophoblasts invasion and to reveal the underlying mechanisms. In the present study, differentially expressed miRs that were screened based on previously published microarray data (GSE96985) and a significantly downregulated miR‑424‑5p (miR‑424) was chosen for further investigation. Subsequently, reverse transcription‑quantitative PCR, CCK‑8, apoptosis, wound healing and Transwell assays were performed to determine the cell viability, apoptotic rate, cell migration and invasion of trophoblast cells. The results showed that miR‑424 was decreased in placenta specimens from patients with PE. Upregulation of miR‑424 promoted cell viability, suppressed cell apoptosis and improved the invasion and migration of trophoblasts, whereas inhibition of miR‑424 had opposite results. Adenomatous polyposis coli (APC), a key mediator of Wnt/β‑catenin signaling pathway, was identified as a functional target of miR‑424 and an inverse relationship was observed between APC and miR‑424 in placenta specimens. Further investigations revealed that APC overexpression efficiently suppressed the effect of miR‑424 in trophoblast cells. In addition, the miR‑424‑mediated effects on trophoblast cells were dependent on the promotion of Wnt/β‑catenin signaling pathway. The present findings revealed that miR‑424 regulates the trophoblast cell invasion by regulating Wnt/β‑catenin pathway through targeting APC, indicating miR‑424 as a potential candidate for the treatment of PE.

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1	Wagner LK: Diagnosis and management of preeclampsia. Am Fam Physician. 70:2317–2324. 2004.PubMed/NCBI
2	Kanasaki K and Kalluri R: The biology of preeclampsia. Kidney Int. 76:831–837. 2009.PubMed/NCBI View Article : Google Scholar
3	Giguère Y, Massé J, Thériault S, Bujold E, Lafond J, Rousseau F and Forest JC: Screening for pre-eclampsia early in pregnancy: Performance of a multivariable model combining clinical characteristics and biochemical markers. BJOG. 122:402–410. 2015.PubMed/NCBI View Article : Google Scholar
4	Ray JG, Vermeulen MJ, Schull MJ and Redelmeier DA: Cardiovascular health after maternal placental syndromes (CHAMPS): Population-based retrospective cohort study. Lancet. 366:1797–1803. 2005.PubMed/NCBI View Article : Google Scholar
5	Meekins JW, Pijnenborg R, Hanssens M, McFadyen IR and van Asshe A: A study of placental bed spiral arteries and trophoblast invasion in normal and severe pre-eclamptic pregnancies. Br J Obstet Gynaecol. 101:669–674. 1994.PubMed/NCBI View Article : Google Scholar
6	Sircar M, Thadhani R and Karumanchi SA: Pathogenesis of preeclampsia. Curr Opin Nephrol Hypertens. 24:131–138. 2015.PubMed/NCBI View Article : Google Scholar
7	Bartel DP: MicroRNAs: Genomics, biogenesis, mechanism, and function. Cell. 116:281–297. 2004.PubMed/NCBI View Article : Google Scholar
8	Takahashi H, Ohkuchi A, Kuwata T, Usui R, Baba Y, Suzuki H, Chaw Kyi TT, Matsubara S, Saito S and Takizawa T: Endogenous and exogenous miR-520c-3p modulates CD44-mediated extravillous trophoblast invasion. Placenta. 50:25–31. 2017.PubMed/NCBI View Article : Google Scholar
9	Fang M, Du H, Han B, Xia G, Shi X, Zhang F, Fu Q and Zhang T: Hypoxia-inducible microRNA-218 inhibits trophoblast invasion by targeting LASP1: Implications for preeclampsia development. Int J Biochem Cell Biol. 87:95–103. 2017.PubMed/NCBI View Article : Google Scholar
10	Liu E, Liu Z, Zhou Y, Chen M, Wang L and Li J: MicroRNA1423p inhibits trophoblast cell migration and invasion by disrupting the TGF-β1/Smad3 signaling pathway. Mol Med Rep. 19:3775–3782. 2019.PubMed/NCBI View Article : Google Scholar
11	Gao Y, She R, Wang Q, Li Y and Zhang H: Up-regulation of miR-299 suppressed the invasion and migration of HTR-8/SVneo trophoblast cells partly via targeting HDAC2 in pre-eclampsia. Biomed Pharmacother. 97:1222–1228. 2018.PubMed/NCBI View Article : Google Scholar
12	Tang Q, Gui J, Wu X and Wu W: Downregulation of miR-424 in placenta is associated with severe preeclampsia. Pregnancy Hypertens. 17:109–112. 2019.PubMed/NCBI View Article : Google Scholar
13	Mouillet JF, Donker RB, Mishima T, Cronqvist T, Chu T and Sadovsky Y: The unique expression and function of miR-424 in human placental trophoblasts. Biol Reprod. 89(25)2013.PubMed/NCBI View Article : Google Scholar
14	Wei S, Li Q, Li Z, Wang L, Zhang L and Xu Z: miR-424-5p promotes proliferation of gastric cancer by targeting Smad3 through TGF-β signaling pathway. Oncotarget. 7:75185–75196. 2016.PubMed/NCBI View Article : Google Scholar
15	Li Y, Liu J, Hu W, Zhang Y, Sang J, Li H, Ma T, Bo Y, Bai T, Guo H, et al: miR-424-5p promotes proliferation, migration and invasion of laryngeal squamous cell carcinoma. Onco Targets Ther. 12:10441–10453. 2019.PubMed/NCBI View Article : Google Scholar
16	Wu K, Hu G, He X, Zhou P, Li J, He B and Sun W: MicroRNA-424-5p suppresses the expression of SOCS6 in pancreatic cancer. Pathol Oncol Res. 19:739–748. 2013.PubMed/NCBI View Article : Google Scholar
17	Abou-Kheir W, Barrak J, Hadadeh O and Daoud G: HTR-8/SVneo cell line contains a mixed population of cells. Placenta. 50:1–7. 2017.PubMed/NCBI View Article : Google Scholar
18	Graham CH, Hawley TS, Hawley RG, MacDougall JR, Kerbel RS, Khoo N and Lala PK: Establishment and characterization of first trimester human trophoblast cells with extended lifespan. Exp Cell Res. 206:204–211. 1993.PubMed/NCBI View Article : Google Scholar
19	Chen D, He B, Zheng P, Wang S, Zhao X, Liu J, Yang X and Cheng W: Identification of mRNA-, circRNA- and lncRNA- associated ceRNA networks and potential biomarkers for preeclampsia from umbilical vein endothelial cells. Front Mol Biosci. 8(652250)2021.PubMed/NCBI View Article : Google Scholar
20	Livak KJ and Schmittgen TD: Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods. 25:402–408. 2001.PubMed/NCBI View Article : Google Scholar
21	Bahar E and Yoon H: Modeling and predicting the cell migration properties from scratch wound healing assay on cisplatin-resistant ovarian cancer cell lines using artificial neural network. Healthcare (Basel). 9(911)2021.PubMed/NCBI View Article : Google Scholar
22	Mao Y, Hou B, Shan L, Sun X and Wang L: Aberrantly up-regulated miR-142-3p inhibited the proliferation and invasion of trophoblast cells by regulating FOXM1. Placenta. 104:253–260. 2021.PubMed/NCBI View Article : Google Scholar
23	Zhou G, Li Z, Hu P, Wang J, Fu J, Wei B and Zhang Y: miR-219a suppresses human trophoblast cell invasion and proliferation by targeting vascular endothelial growth factor receptor 2 (VEGFR2). J Assist Reprod Genet. 38:461–470. 2021.PubMed/NCBI View Article : Google Scholar
24	Wang Y, Du X and Wang J: Transfer of miR-15a-5p by placental exosomes promotes pre-eclampsia progression by regulating PI3K/AKT signaling pathway via CDK1. Mol Immunol. 128:277–286. 2020.PubMed/NCBI View Article : Google Scholar
25	Wang YP, Zhao P, Liu JY, Liu SM and Wang YX: MicroRNA-132 stimulates the growth and invasiveness of trophoblasts by targeting DAPK-1. Eur Rev Med Pharmacol Sci. 24:9837–9843. 2020.PubMed/NCBI View Article : Google Scholar
26	Han L, Luo QQ, Peng MG, Zhang Y and Zhu XH: miR-483 is downregulated in pre-eclampsia via targeting insulin-like growth factor 1 (IGF1) and regulates the PI3K/Akt/mTOR pathway of endothelial progenitor cells. J Obstet Gynaecol Res. 47:63–72. 2021.PubMed/NCBI View Article : Google Scholar
27	Lv Y, Lu X, Li C, Fan Y, Ji X, Long W, Meng L, Wu L, Wang L, Lv M and Ding H: miR-145-5p promotes trophoblast cell growth and invasion by targeting FLT1. Life Sci. 239(117008)2019.PubMed/NCBI View Article : Google Scholar
28	Li X, Li C, Wang Y, Cai J, Zhao L, Su Z and Ye H: IGFBP1 inhibits the invasion, migration, and apoptosis of HTR-8/SVneo trophoblast cells in preeclampsia. Hypertens Pregnancy. 41:53–63. 2022.PubMed/NCBI View Article : Google Scholar
29	Yu Y, An X and Fan D: Histone deacetylase sirtuin 2 enhances viability of trophoblasts through p65-mediated microRNA-146a/ACKR2 axis. Reprod Sci. 28:1370–1381. 2021.PubMed/NCBI View Article : Google Scholar
30	Li L, Peng W, Zhou Q, Wan JP, Wang XT and Qi HB: LRP6 regulates Rab7-mediated autophagy through the Wnt/β-catenin pathway to modulate trophoblast cell migration and invasion. J Cell Biochem. 121:1599–1609. 2020.PubMed/NCBI View Article : Google Scholar
31	Douchi D, Ohtsuka H, Ariake K, Masuda K, Kawasaki S, Kawaguchi K, Fukase K, Oikawa M, Motoi F, Naitoh T, et al: Silencing of LRRFIP1 reverses the epithelial-mesenchymal transition via inhibition of the Wnt/β-catenin signaling pathway. Cancer Lett. 365:132–140. 2015.PubMed/NCBI View Article : Google Scholar
32	Clevers H and Nusse R: Wnt/β-catenin signaling and disease. Cell. 149:1192–1205. 2012.PubMed/NCBI View Article : Google Scholar
33	Anastas JN and Moon RT: WNT signalling pathways as therapeutic targets in cancer. Nat Rev Cancer. 13:11–26. 2013.PubMed/NCBI View Article : Google Scholar
34	Carter AM, Enders AC and Pijnenborg R: The role of invasive trophoblast in implantation and placentation of primates. Philos Trans R Soc Lond B Biol Sci. 370(20140070)2015.PubMed/NCBI View Article : Google Scholar
35	Jiang L, Long A, Tan L, Hong M, Wu J, Cai L and Li Q: Elevated microRNA-520g in pre-eclampsia inhibits migration and invasion of trophoblasts. Placenta. 51:70–75. 2017.PubMed/NCBI View Article : Google Scholar
36	Xiao J, Tao T, Yin Y, Zhao L, Yang L and Hu L: miR-144 may regulate the proliferation, migration and invasion of trophoblastic cells through targeting PTEN in preeclampsia. Biomed Pharmacother. 94:341–353. 2017.PubMed/NCBI View Article : Google Scholar
37	Lu M, Kong X, Wang H, Huang G, Ye C and He Z: A novel microRNAs expression signature for hepatocellular carcinoma diagnosis and prognosis. Oncotarget. 8:8775–8784. 2017.PubMed/NCBI View Article : Google Scholar
38	Liu X, Fu Y, Zhang G, Zhang D, Liang N, Li F, Li C, Sui C, Jiang J, Lu H, et al: miR-424-5p promotes anoikis resistance and lung metastasis by inactivating hippo signaling in thyroid cancer. Mol Ther Oncolytics. 15:248–260. 2019.PubMed/NCBI View Article : Google Scholar
39	Wang J, Wang S, Zhou J and Qian Q: miR-424-5p regulates cell proliferation, migration and invasion by targeting doublecortin-like kinase 1 in basal-like breast cancer. Biomed Pharmacother. 102:147–152. 2018.PubMed/NCBI View Article : Google Scholar
40	Li C and Li Q: Circular RNA circ_0111277 serves as ceRNA, targeting the miR-424-5p/NFAT5 axis to regulate the proliferation, migration, and invasion of trophoblast cells in preeclampsia. Reprod Sci. 29:923–935. 2022.PubMed/NCBI View Article : Google Scholar
41	Wang X, Zhang Z, Zeng X, Wang J, Zhang L, Song W and Shi Y: Wnt/β-catenin signaling pathway in severe preeclampsia. J Mol Histol. 49:317–327. 2018.PubMed/NCBI View Article : Google Scholar
42	Fitzgerald JS, Germeyer A, Huppertz B, Jeschke U, Knöfler M, Moser G, Scholz C, Sonderegger S, Toth B and Markert UR: Governing the invasive trophoblast: Current aspects on intra- and extracellular regulation. Am J Reprod Immunol. 63:492–505. 2010.PubMed/NCBI View Article : Google Scholar
43	Zhang Z, Wang X, Zhang L, Shi Y, Wang J and Yan H: Wnt/β-catenin signaling pathway in trophoblasts and abnormal activation in preeclampsia (review). Mol Med Rep. 16:1007–1013. 2017.PubMed/NCBI View Article : Google Scholar
44	Tian FJ, Qin CM, Li XC, Wu F, Liu XR, Xu WM and Lin Y: Decreased stathmin-1 expression inhibits trophoblast proliferation and invasion and is associated with recurrent miscarriage. Am J Pathol. 185:2709–2721. 2015.PubMed/NCBI View Article : Google Scholar
45	Xing Y, Clements WK, Le Trong I, Hinds TR, Stenkamp R, Kimelman D and Xu W: Crystal structure of a beta-catenin/APC complex reveals a critical role for APC phosphorylation in APC function. Mol Cell. 15:523–533. 2004.PubMed/NCBI View Article : Google Scholar
46	Sahay AS, Jadhav AT, Sundrani DP, Wagh GN, Mehendale SS and Joshi SR: Matrix metalloproteinases-2 (MMP-2) and matrix metalloproteinases-9 (MMP-9) are differentially expressed in different regions of normal and preeclampsia placentae. J Cell Biochem. 119:6657–6664. 2018.PubMed/NCBI View Article : Google Scholar
47	Li Z, Takino T, Endo Y and Sato H: Activation of MMP-9 by membrane type-1 MMP/MMP-2 axis stimulates tumor metastasis. Cancer Sci. 108:347–353. 2017.PubMed/NCBI View Article : Google Scholar
48	Dastmalchi N, Hosseinpourfeizi MA, Khojasteh SMB, Baradaran B and Safaralizadeh R: Tumor suppressive activity of miR-424-5p in breast cancer cells through targeting PD-L1 and modulating PTEN/PI3K/AKT/mTOR signaling pathway. Life Sci. 259(118239)2020.PubMed/NCBI View Article : Google Scholar
49	Dastmalchi N, Safaralizadeh R, Hosseinpourfeizi MA, Baradaran B and Khojasteh SMB: MicroRNA-424-5p enhances chemosensitivity of breast cancer cells to Taxol and regulates cell cycle, apoptosis, and proliferation. Mol Biol Rep. 48:1345–1357. 2021.PubMed/NCBI View Article : Google Scholar
50	Zou Z, He Z, Cai J, Huang L, Zhu H and Luo Y: Potential role of microRNA-424 in regulating ERRgamma to suppress trophoblast proliferation and invasion in fetal growth restriction. Placenta. 83:57–62. 2019.PubMed/NCBI View Article : Google Scholar
51	Liu C, Li H, Zhang Y and Ding H: Long intergenic noncoding RNA 00473 promoting migration and invasion of trophoblastic cell line HTR-8/SVneo via regulating miR-424-5p-mediated wnt3a/β-catenin signaling pathway. J Obstet Gynaecol Res. 47:3034–3046. 2021.PubMed/NCBI View Article : Google Scholar
52	Selvaraj S, Lakshmanan G, Kalimuthu K and Sekar D: Role of microRNAs and their involvement in preeclampsia. Epigenomics. 12:1765–1767. 2020.PubMed/NCBI View Article : Google Scholar