MicroRNA‑155‑5p inhibits trophoblast cell proliferation and invasion by disrupting centrosomal function

Tsai,Yung-Chieh; Kuo,Tian-Ni; Lin,Ruei-Ci; Tsai,Hui-Ling; Chao,Yu-Ying; Lee,Pei-Rong; Su,Ping-Jui; Wang,Chia-Yih

doi:10.3892/mmr.2024.13209

MicroRNA‑155‑5p inhibits trophoblast cell proliferation and invasion by disrupting centrosomal function

Authors:
- Yung-Chieh Tsai
- Tian-Ni Kuo
- Ruei-Ci Lin
- Hui-Ling Tsai
- Yu-Ying Chao
- Pei-Rong Lee
- Ping-Jui Su
- Chia-Yih Wang
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Affiliations: Department of Obstetrics and Gynecology, Chi‑Mei Medical Center, Tainan 710, Taiwan, R.O.C., Institute of Basic Medical Sciences, College of Medicine, National Cheng Kung University, Tainan 701, Taiwan, R.O.C., Department of Cell Biology and Anatomy, College of Medicine, National Cheng Kung University, Tainan 701, Taiwan, R.O.C., Division of General Surgery, Department of Surgery, National Cheng Kung University Hospital, Tainan 704, Taiwan, R.O.C.
Published online on: March 22, 2024 https://doi.org/10.3892/mmr.2024.13209
Article Number: 85
Copyright: © Tsai et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Recurrent miscarriage is used to refer to more than three pregnancy failures before 20 weeks of gestation. Defective trophoblast cell growth and invasion are frequently observed in recurrent miscarriage. Several microRNAs (miRs), including miR‑155‑5p, are aberrantly upregulated in recurrent miscarriage; however, the underlying molecular mechanisms remain unclear. The centrosome orchestrates microtubule networks and coordinates cell cycle progression. In addition, it is a base for primary cilia, which are antenna‑like organelles that coordinate signaling during development and growth. Thus, deficiencies in centrosomal functions can lead to several disease, such as breast cancer and microcephaly. In the present study, the signaling cascades were analyzed by western blotting, and the centrosome and primary cilia were observed and analyzed by immunofluorescence staining. The results showed that overexpression of miR‑155‑5p induced centrosome amplification and blocked primary cilia formation in trophoblast cells. Notably, centrosome amplification inhibited trophoblast cell growth by upregulating apoptotic cleaved‑caspase 3 and cleaved‑poly (ADP‑ribose) polymerase in miR‑155‑5p‑overexpressing trophoblast cells. In addition, overexpression of miR‑155‑5p inhibited primary cilia formation, thereby inhibiting epithelial‑mesenchymal transition and trophoblast cell invasion. All phenotypes could be rescued when cells were co‑transfected with the miR‑155‑5p inhibitor, thus supporting the role of miR‑155‑5p in centrosomal functions. It was also found that miR‑155‑5p activated autophagy, whereas disruption of autophagy via the depletion of autophagy‑related 16‑like 1 alleviated miR‑155‑5p‑induced apoptosis and restored trophoblast cell invasion. In conclusion, the present study indicated a novel role of miR‑55‑5p in mediating centrosomal function in recurrent miscarriage.

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1	Bashiri A, Halper KI and Orvieto R: Recurrent implantation failure-update overview on etiology, diagnosis, treatment and future directions. Reprod Biol Endocrinol. 16:1212018. View Article : Google Scholar : PubMed/NCBI
2	Ford HB and Schust DJ: Recurrent pregnancy loss: Etiology, diagnosis, and therapy. Rev Obstet Gynecol. 2:76–83. 2009.PubMed/NCBI
3	Rai R and Regan L: Recurrent miscarriage. Lancet. 368:601–611. 2006. View Article : Google Scholar : PubMed/NCBI
4	You Y, Stelzl P, Joseph DN, Aldo PB, Maxwell AJ, Dekel N, Liao A, Whirledge S and Mor G: TNF-α regulated endometrial stroma secretome promotes trophoblast invasion. Front Immunol. 12:7374012021. View Article : Google Scholar : PubMed/NCBI
5	Zhang S, Lin H, Kong S, Wang S, Wang H, Wang H and Armant DR: Physiological and molecular determinants of embryo implantation. Mol Aspects Med. 34:939–980. 2013. View Article : Google Scholar : PubMed/NCBI
6	Pan-Castillo B, Gazze SA, Thomas S, Lucas C, Margarit L, Gonzalez D, Francis LW and Conlan RS: Morphophysical dynamics of human endometrial cells during decidualization. Nanomedicine. 14:2235–2245. 2018. View Article : Google Scholar : PubMed/NCBI
7	Vento-Tormo R, Efremova M, Botting RA, Turco MY, Vento-Tormo M, Meyer KB, Park JE, Stephenson E, Polański K, Goncalves A, et al: Single-cell reconstruction of the early maternal-fetal interface in humans. Nature. 563:347–353. 2018. View Article : Google Scholar : PubMed/NCBI
8	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. 370:201400702015. View Article : Google Scholar : PubMed/NCBI
9	Meakin C, Barrett ES and Aleksunes LM: Extravillous trophoblast migration and invasion: Impact of environmental chemicals and pharmaceuticals. Reprod Toxicol. 107:60–68. 2022. View Article : Google Scholar : PubMed/NCBI
10	Ambros V: The functions of animal microRNAs. Nature. 431:350–355. 2004. View Article : Google Scholar : PubMed/NCBI
11	Jouravleva K, Golovenko D, Demo G, Dutcher RC, Hall TMT, Zamore PD and Korostelev AA: Structural basis of microRNA biogenesis by Dicer-1 and its partner protein Loqs-PB. Mol Cell. 82:4049–4063.e6. 2022. View Article : Google Scholar : PubMed/NCBI
12	Basavarajappa D, Uebbing S, Kreiss M, Lukic A, Suess B, Steinhilber D, Samuelsson B and Rådmark O: Dicer up-regulation by inhibition of specific proteolysis in differentiating monocytic cells. Proc Natl Acad Sci USA. 117:8573–8583. 2020. View Article : Google Scholar : PubMed/NCBI
13	Muys BR, Sousa JF, Plaça JR, de Araújo LF, Sarshad AA, Anastasakis DG, Wang X, Li XL, de Molfetta GA, Ramão A, et al: miR-450a acts as a tumor suppressor in ovarian cancer by regulating energy metabolism. Cancer Res. 79:3294–3305. 2019. View Article : Google Scholar : PubMed/NCBI
14	Singh R, Yadav V, Kumar S and Saini N: MicroRNA-195 inhibits proliferation, invasion and metastasis in breast cancer cells by targeting FASN, HMGCR, ACACA and CYP27B1. Sci Rep. 5:174542015. View Article : Google Scholar : PubMed/NCBI
15	Sun Z, Shi K, Yang S, Liu J, Zhou Q, Wang G, Song J, Li Z, Zhang Z and Yuan W: Effect of exosomal miRNA on cancer biology and clinical applications. Mol Cancer. 17:1472018. View Article : Google Scholar : PubMed/NCBI
16	Skalis G, Katsi V, Miliou A, Georgiopoulos G, Papazachou O, Vamvakou G, Nihoyannopoulos P, Tousoulis D and Makris T: MicroRNAs in preeclampsia. Microrna. 8:28–35. 2019. View Article : Google Scholar : PubMed/NCBI
17	Zhang H, He Y, Wang JX, Chen MH, Xu JJ, Jiang MH, Feng YL and Gu YF: miR-30-5p-mediated ferroptosis of trophoblasts is implicated in the pathogenesis of preeclampsia. Redox Biol. 29:1014022020. View Article : Google Scholar : PubMed/NCBI
18	Wang CY, Tsai PY, Chen TY, Tsai HL, Kuo PL and Su MT: Elevated miR-200a and miR-141 inhibit endocrine gland-derived vascular endothelial growth factor expression and ciliogenesis in preeclampsia. J Physiol. 597:3069–3083. 2019. View Article : Google Scholar : PubMed/NCBI
19	Chen CH, Lu F, Yang WJ, Yang PE, Chen WM, Kang ST, Huang YS, Kao YC, Feng CT, Chang PC, et al: A novel platform for discovery of differentially expressed microRNAs in patients with repeated implantation failure. Fertil Steril. 116:181–188. 2021. View Article : Google Scholar : PubMed/NCBI
20	Lai PY, Wang CY, Chen WY, Kao YH, Tsai HM, Tachibana T, Chang WC and Chung BC: Steroidogenic factor 1 (NR5A1) resides in centrosomes and maintains genomic stability by controlling centrosome homeostasis. Cell Death Differ. 18:1836–1844. 2011. View Article : Google Scholar : PubMed/NCBI
21	Chen TY, Lien WC, Cheng HL, Kuan TS, Sheu SY and Wang CY: Chloroquine inhibits human retina pigmented epithelial cell growth and microtubule nucleation by downregulating p150^glued. J Cell Physiol. 234:10445–10457. 2019. View Article : Google Scholar : PubMed/NCBI
22	Lin RC, Chao YY, Lien WC, Chang HC, Tsai SW and Wang CY: Polo-like kinase 1 selective inhibitor BI2536 (dihydropteridinone) disrupts centrosome homeostasis via ATM-ERK cascade in adrenocortical carcinoma. Oncol Rep. 50:1672023. View Article : Google Scholar : PubMed/NCBI
23	Ferguson RL and Maller JL: Centrosomal localization of cyclin E-Cdk2 is required for initiation of DNA synthesis. Curr Biol. 20:856–860. 2010. View Article : Google Scholar : PubMed/NCBI
24	Chen TY, Lin TC, Kuo PL, Chen ZR, Cheng HL, Chao YY, Syu JS, Lu FI and Wang CY: Septin 7 is a centrosomal protein that ensures S phase entry and microtubule nucleation by maintaining the abundance of p150^glued. J Cell Physiol. 236:2706–2724. 2021. View Article : Google Scholar : PubMed/NCBI
25	Chao YY, Huang BM, Peng IC, Lee PR, Lai YS, Chiu WT, Lin YS, Lin SC, Chang JH, Chen PS, et al: ATM- and ATR-induced primary ciliogenesis promotes cisplatin resistance in pancreatic ductal adenocarcinoma. J Cell Physiol. 237:4487–4503. 2022. View Article : Google Scholar : PubMed/NCBI
26	Hildebrandt F, Benzing T and Katsanis N: Ciliopathies. N Engl J Med. 364:1533–1543. 2011. View Article : Google Scholar : PubMed/NCBI
27	Yamamoto Y and Mizushima N: Autophagy and ciliogenesis. JMA J. 4:207–215. 2021. View Article : Google Scholar : PubMed/NCBI
28	Truong ME, Bilekova S, Choksi SP, Li W, Bugaj LJ, Xu K and Reiter JF: Vertebrate cells differentially interpret ciliary and extraciliary cAMP. Cell. 184:2911–2926.e18. 2021. View Article : Google Scholar : PubMed/NCBI
29	Tsai YC, Kuo TN, Chao YY, Lee PR, Lin RC, Xiao XY, Huang BM and Wang CY: PDGF-AA activates AKT and ERK signaling for testicular interstitial Leydig cell growth via primary cilia. J Cell Biochem. 124:89–102. 2023. View Article : Google Scholar : PubMed/NCBI
30	Wang CY, Tsai HL, Syu JS, Chen TY and Su MT: Primary cilium-regulated EG-VEGF signaling facilitates trophoblast invasion. J Cell Physiol. 232:1467–1477. 2017. View Article : Google Scholar : PubMed/NCBI
31	Gu X, Liu H, Luo W, Wang X, Wang H and Li L: Di-2-ethylhexyl phthalate-induced miR-155-5p promoted lipid metabolism via inhibiting cAMP/PKA signaling pathway in human trophoblastic HTR-8/Svneo cells. Reprod Toxicol. 114:22–31. 2022. View Article : Google Scholar : PubMed/NCBI
32	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. View Article : Google Scholar : PubMed/NCBI
33	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. View Article : Google Scholar : PubMed/NCBI
34	Tan HX, Yang SL, Li MQ and Wang HY: Autophagy suppression of trophoblast cells induces pregnancy loss by activating decidual NK cytotoxicity and inhibiting trophoblast invasion. Cell Commun Signal. 18:732020. View Article : Google Scholar : PubMed/NCBI
35	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. View Article : Google Scholar : PubMed/NCBI
36	Orth JD, Loewer A, Lahav G and Mitchison TJ: Prolonged mitotic arrest triggers partial activation of apoptosis, resulting in DNA damage and p53 induction. Mol Biol Cell. 23:567–576. 2012. View Article : Google Scholar : PubMed/NCBI
37	Wang CY, Kao YH, Lai PY, Chen WY and Chung BC: Steroidogenic factor 1 (NR5A1) maintains centrosome homeostasis in steroidogenic cells by restricting centrosomal DNA-dependent protein kinase activation. Mol Cell Biol. 33:476–484. 2013. View Article : Google Scholar : PubMed/NCBI
38	Bahmanyar S, Guiney EL, Hatch EM, Nelson WJ and Barth AIM: Formation of extra centrosomal structures is dependent on beta-catenin. J Cell Sci. 123:3125–3135. 2010. View Article : Google Scholar : PubMed/NCBI
39	Wang CY, Lai PY, Chen TY and Chung BC: NR5A1 prevents centriole splitting by inhibiting centrosomal DNA-PK activation and β-catenin accumulation. Cell Commun Signal. 12:552014. View Article : Google Scholar : PubMed/NCBI
40	Illsley NP, DaSilva-Arnold SC, Zamudio S, Alvarez M and Al-Khan A: Trophoblast invasion: Lessons from abnormally invasive placenta (placenta accreta). Placenta. 102:61–66. 2020. View Article : Google Scholar : PubMed/NCBI
41	Ming Z, Vining B, Bagheri-Fam S and Harley V: SOX9 in organogenesis: Shared and unique transcriptional functions. Cell Mol Life Sci. 79:5222022. View Article : Google Scholar : PubMed/NCBI
42	Edelman HE, McClymont SA, Tucker TR, Pineda S, Beer RL, McCallion AS and Parsons MJ: SOX9 modulates cancer biomarker and cilia genes in pancreatic cancer. Hum Mol Genet. 30:485–499. 2021. View Article : Google Scholar : PubMed/NCBI
43	Xu WP, Cui YL, Chen LL, Ding K, Ding CH, Chen F, Zhang X and Xie WF: Deletion of Sox9 in the liver leads to hepatic cystogenesis in mice by transcriptionally downregulating Sec63. J Pathol. 254:57–69. 2021.PubMed/NCBI
44	Schepers GE, Teasdale RD and Koopman P: Twenty pairs of sox: Extent, homology, and nomenclature of the mouse and human sox transcription factor gene families. Dev Cell. 3:167–170. 2002. View Article : Google Scholar : PubMed/NCBI
45	Tew SR and Hardingham TE: Regulation of SOX9 mRNA in human articular chondrocytes involving p38 MAPK activation and mRNA stabilization. J Biol Chem. 281:39471–39479. 2006. View Article : Google Scholar : PubMed/NCBI
46	Bao Z, Zhang N, Niu W, Mu M, Zhang X, Hu S and Niu C: Exosomal miR-155-5p derived from glioma stem-like cells promotes mesenchymal transition via targeting ACOT12. Cell Death Dis. 13:7252022. View Article : Google Scholar : PubMed/NCBI
47	Chen G, Wang D, Zhao X, Cao J, Zhao Y, Wang F, Bai J, Luo D and Li L: miR-155-5p modulates malignant behaviors of hepatocellular carcinoma by directly targeting CTHRC1 and indirectly regulating GSK-3β-involved Wnt/β-catenin signaling. Cancer Cell Int. 17:1182017. View Article : Google Scholar : PubMed/NCBI
48	Yang LW, Wu XJ, Liang Y, Ye GQ, Che YC, Wu XZ, Zhu XJ, Fan HL, Fan XP and Xu JF: miR-155 increases stemness and decitabine resistance in triple-negative breast cancer cells by inhibiting TSPAN5. Mol Carcinog. 59:447–461. 2020. View Article : Google Scholar : PubMed/NCBI
49	Wu HY, Liu K and Zhang JL: LINC00240/miR-155 axis regulates function of trophoblasts and M2 macrophage polarization via modulating oxidative stress-induced pyroptosis in preeclampsia. Mol Med. 28:1192022. View Article : Google Scholar : PubMed/NCBI