Abstract
Preeclampsia is the main reason for maternal and fetal deaths during the second half of pregnancy. Trophoblast cells play a pivotal role in preeclampsia progression. Circular RNA (circRNA) circ_0111277 has been reported to be related to the development of trophoblast cells. This study is designed to explore the role and mechanism of circ_0111277 on trophoblast cell behavior in preeclampsia. Circ_0111277, microRNA-424-5p (miR-424-5p), and nuclear factor of activated T-cell 5 (NFAT5) levels were measured by real-time quantitative polymerase chain reaction (RT-qPCR). Cell viability, migration, invasion, and angiogenesis were measured by 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay, transwell assay, tube formation assay, and wound healing assay. Protein levels of matrix metallopeptidase 2 (MMP2), vascular endothelial growth factor-A (VEGF-A), NFAT5, phospho-phosphatidylinositol 3 kinase (p-PI3K), PI3K, phospho-protein kinase B (p-AKT), and AKT were examined by western blot assay. The binding relationship between miR-424-5p and circ_0111277 or NFAT5 was predicted by circBank or starBase and then verified by a dual-luciferase reporter assay. Circ_0111277 and NFAT5 expression were increased in placenta tissues of preeclampsia patients, and miR-424-5p was decreased. Moreover, circ_0111277 knockdown could boost cell viability, migration, invasion, and angiogenesis in trophoblast cells. The mechanical analysis discovered that circ_0111277 acted as a sponge of miR-424-5p to regulate NFAT5 expression. Besides, circ_0111277 silencing promoted the PI3K/AKT signaling pathway in trophoblast cells. Circ_0111277 downregulation could facilitate cell growth and metastasis in trophoblast cells partly by regulating the miR-424-5p/NFAT5 axis, providing an underlying circRNA-targeted therapy for preeclampsia.
Similar content being viewed by others
References
Arroyo JA, Teng C, Battaglia FC, Galan HL. Determination of the NFAT5/TonEBP transcription factor in the human and ovine placenta. Syst Biol Reprod Med. 2009;55(4):164–70.
Bai Y, Rao H, Chen W, Luo X, Tong C, Qi H. Profiles of circular RNAs in human placenta and their potential roles related to preeclampsia. Biol Reprod. 2018;98(5):705–12.
Chen J, Yue C, Xu J, Zhan Y, Zhao H, Li Y, Ye Y. Downregulation of receptor tyrosine kinase-like orphan receptor 1 in preeclampsia placenta inhibits human trophoblast cell proliferation, migration, and invasion by PI3K/AKT/mTOR pathway accommodation. Placenta. 2019;82:17–24.
Deng N, Lei D, Huang J, Yang Z, Fan C, Wang S. Circular RNA expression profiling identifies hsa_circ_0011460 as a novel molecule in severe preeclampsia. Pregnancy Hypertens. 2019;17:216–25.
Filipek A, Jurewicz E. Preeclampsia - a disease of pregnant women. Postepy Biochem. 2018;64(4):232–29.
Ghosh R, Lipson KL, Sargent KE, Mercurio AM, Hunt JS, Ron D, Urano F. Transcriptional regulation of VEGF-A by the unfolded protein response pathway. PLoS One. 2010;5(3):e9575.
Guo JU, Agarwal V, Guo H, Bartel DP. Expanded identification and characterization of mammalian circular RNAs. Genome Biol. 2014;15(7):409.
Hansen TB, Jensen TI, Clausen BH, Bramsen JB, Finsen B, Damgaard CK, Kjems J. Natural RNA circles function as efficient microRNA sponges. Nature. 2013;495(7441):384–8.
Hansen YB, Myrhøj V, Jørgensen FS, Oxvig C, Sørensen S. First trimester PAPP-A2, PAPP-A and hCGβ in small-for-gestational-age pregnancies. Clin Chem Lab Med. 2016;54(1):117–23.
Jia N, Li J. Role of circular RNAs in preeclampsia. Dis Markers. 2019;2019:7237495.
Kagami H, Akutsu T, Maegawa S, Hosokawa H, Nacher JC. Determining associations between Human Diseases and non-coding RNAs with Critical Roles in Network Control. Sci Rep. 2015;5:14577.
Kristensen LS, Andersen MS, Stagsted LVW, Ebbesen KK, Hansen TB, Kjems J. The biogenesis, biology and characterization of circular RNAs. Nat Rev Genet. 2019;20(11):675–91.
Kulcheski FR, Christoff AP, Margis R. Circular RNAs are miRNA sponges and can be used as a new class of biomarker. J Biotechnol. 2016;238:42–51.
Lala PK, Chakraborty C. Factors regulating trophoblast migration and invasiveness: possible derangements contributing to pre-eclampsia and fetal injury. Placenta. 2003;24(6):575–87.
Lee CQ, Gardner L, Turco M, Zhao N, Murray MJ, Coleman N, Rossant J, Hemberger M, Moffett A. What is trophoblast? A combination of criteria define human first-trimester trophoblast. Stem Cell Reports. 2016;6(2):257–72.
Lee GT, Price MD, Mejia CA, Galan HL, Arroyo JA. Increased trophoblast expression of NFAT5/TonEBP in pre-eclamptic placentas and hyperosmolar-treated BeWo cells. Eur J Obstet Gynecol Reprod Biol. 2014;183:37–43.
Li X, Yang R, Xu Y, Zhang Y. Circ_0001438 participates in the pathogenesis of preeclampsia via the circ_0001438/miR-942/NLRP3 regulatory network. Placenta. 2020;104:40–50.
Liao HQ, Liu H, Sun HL, Xiang JB, Wang XX, Jiang CX, Ma L, Cao ZG. MiR-361-3p/Nfat5 signaling axis controls cementoblast Differentiation. J Dent Res. 2019;98(10):1131–9.
Liu S, Xie X, Lei H, Zou B, Xie L. Identification of Key circRNAs/lncRNAs/miRNAs/mRNAs and pathways in preeclampsia using bioinformatics Analysis. Med Sci Monit. 2019;25:1679–93.
López-Jiménez E, Rojas AM, Andrés-León E. RNA sequencing and prediction tools for circular RNAs analysis. Adv Exp Med Biol. 2018;1087:17–33.
Macintire K, Tuohey L, Ye L, Palmer K, Gantier M, Tong S, Kaitu'u-Lino TJ. PAPPA2 is increased in severe early onset pre-eclampsia and upregulated with hypoxia. Reprod Fertil Dev. 2014;26(2):351–7.
Memczak S, Jens M, Elefsinioti A, Torti F, Krueger J, Rybak A, Maier L, Mackowiak SD, Gregersen LH, Munschauer M, Loewer A, Ziebold U, Landthaler M, Kocks C, le Noble F, Rajewsky N. Circular RNAs are a large class of animal RNAs with regulatory potency. Nature. 2013;495(7441):333–8.
Mouillet JF, Donker RB, Mishima T, Cronqvist T, Chu T, Sadovsky Y. The unique expression and function of miR-424 in human placental trophoblasts. Biol Reprod. 2013;89(2):25.
Onogi A, Naruse K, Sado T, Tsunemi T, Shigetomi H, Noguchi T, Yamada Y, Akasaki M, Oi H, Kobayashi H. Hypoxia inhibits invasion of extravillous trophoblast cells through reduction of matrix metalloproteinase (MMP)-2 activation in the early first trimester of human pregnancy. Placenta. 2011;32(9):665–70.
Ou Y, Zhu L, Wei X, Bai S, Chen M, Chen H, Zhang J. Circular RNA circ_0111277 attenuates human trophoblast cell invasion and migration by regulating miR-494/HTRA1/Notch-1 signal pathway in pre-eclampsia. Cell Death Dis. 2020;11(6):479.
Panda AC. Circular RNAs Act as miRNA Sponges. Adv Exp Med Biol. 2018;1087:67–79.
Park JK, Kang TG, Kang MY, Park JE, Cho IA, Shin JK, Choi WJ, Lee SA, Choi WS, Kwon HM, Lee JH, Paik WY. Increased NFAT5 expression stimulates transcription of Hsp70 in preeclamptic placentas. Placenta. 2014;35(2):109–16.
Patop IL, Wüst S, Kadener S. Past, present, and future of circRNAs. Embo j. 2019;38(16):e100836.
Phipps E, Prasanna D, Brima W, Jim B. Preeclampsia: updates in pathogenesis, definitions, and guidelines. Clin J Am Soc Nephrol. 2016;11(6):1102–13.
Qian Y, Lu Y, Rui C, Qian Y, Cai M, Jia R. Potential significance of circular RNA in human placental tissue for patients with preeclampsia. Cell Physiol Biochem. 2016;39(4):1380–90.
Rana S, Lemoine E, Granger JP, Karumanchi SA. Preeclampsia: pathophysiology, challenges, and perspectives. Circ Res. 2019;124(7):1094–112.
Rong D, Sun H, Li Z, Liu S, Dong C, Fu K, Tang W, Cao H. An emerging function of circRNA-miRNAs-mRNA axis in human diseases. Oncotarget. 2017;8(42):73271–81.
Sekar S, Geiger P, Cuyugan L, Boyle A, Serrano G, Beach TG, Liang WS. Identification of circular RNAs using RNA sequencing. J Vis Exp. 2019;153.
Shafabakhsh R, Mirhosseini N, Chaichian S, Moazzami B, Mahdizadeh Z, Asemi Z. Could circRNA be a new biomarker for pre-eclampsia? Mol Reprod Dev. 2019;86(12):1773–80.
Tang Q, Gui J, Wu X, Wu W. Downregulation of miR-424 in placenta is associated with severe preeclampsia. Pregnancy Hypertens. 2019;17:109–12.
Tang R, Zhang Z, Han W. CircLRRK1 targets miR-223-3p to inhibit the proliferation, migration and invasion of trophoblast cells by regulating the PI3K/AKT signaling pathway. Placenta. 2020;104:110–8.
Thomas LF, Sætrom P. Circular RNAs are depleted of polymorphisms at microRNA binding sites. Bioinformatics. 2014;30(16):2243–6.
Velicky P, Windsperger K, Petroczi K, Pils S, Reiter B, Weiss T, Vondra S, Ristl R, Dekan S, Fiala C, Cantonwine DE, McElrath TF, Jilma B, Knöfler M, Boehm T, Pollheimer J. Pregnancy-associated diamine oxidase originates from extravillous trophoblasts and is decreased in early-onset preeclampsia. Sci Rep. 2018;8(1):6342.
Zhang H, Hou L, Li CM, Zhang WY. The chemokine CXCL6 restricts human trophoblast cell migration and invasion by suppressing MMP-2 activity in the first trimester. Hum Reprod. 2013;28(9):2350–62.
Zhang Y., Yang H., Zhang Y., Shi J., Chen R. circCRAMP1L is a novel biomarker of preeclampsia risk and may play a role in preeclampsia pathogenesis via regulation of the MSP/RON axis in trophoblasts. BMC Pregnancy Childbirth, 2020; 20(1):652.
Zhang Y, Yang H, Zhang Y, Shi J, Chen R, Xiao X. CircSFXN1 regulates the behaviour of trophoblasts and likely mediates preeclampsia. Placenta. 2020;101:115–23.
Zhou B, Zhang X, Li T, Xie R, Zhou J, Luo Y, Yang C. CircZDHHC20 represses the proliferation, migration and invasion in trophoblast cells by miR-144/GRHL2 axis. Cancer Cell Int. 2020;20:19.
Availability of Data and Materials
Please contact the correspondence author for the data request.
Author information
Authors and Affiliations
Contributions
Chunhua Li was responsible for drafting the manuscript. Chunhua Li and Qing Li contributed to the analysis and interpretation of data. Chunhua Li and Qing Li contributed in the data collection. All authors read and approved the final manuscript.
Corresponding author
Ethics declarations
Ethics Approval and Informed Consent
Written informed consent was obtained from patients with approval by the Institutional Review Board in Qingpu Branch of Zhongshan Hospital Affiliated to Fudan University.
Consent to Participate
Not applicable
Consent for Publication
Not applicable
Conflict of Interest
The authors declare no competing interests.
Additional information
Publisher’s Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Supplementary Information
ESM 1
(PNG 44.9 kb)
Rights and permissions
About this article
Cite this article
Li, C., 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). https://doi.org/10.1007/s43032-021-00715-y
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s43032-021-00715-y