Downregulation of lncRNA <em>USP2‑AS1</em> in the placentas of pregnant women with non‑diabetic fetal macrosomia promotes trophoblast cell proliferation

Lu,Yiwen; Tang,Qiuqin; Yang,Shanshan; Cheng,Yuting; Li,Mei; Guo,Dan; Fu,Ziqiang; Jiang,Hua; Wu,Wei

doi:10.3892/mmr.2022.12766

Downregulation of lncRNA USP2‑AS1 in the placentas of pregnant women with non‑diabetic fetal macrosomia promotes trophoblast cell proliferation

Authors:
- Yiwen Lu
- Qiuqin Tang
- Shanshan Yang
- Yuting Cheng
- Mei Li
- Dan Guo
- Ziqiang Fu
- Hua Jiang
- Wei Wu
View Affiliations

Affiliations: State Key Laboratory of Reproductive Medicine, Institute of Toxicology, Nanjing Medical University, Nanjing, Jiangsu 211166, P.R. China, Department of Gynecology and Obstetrics, Women's Hospital of Nanjing Medical University, Nanjing Maternity and Child Health Care Hospital, Nanjing, Jiangsu 210004, P.R. China, School of Public Health, Nanjing Medical University, Nanjing, Jiangsu 211100, P.R. China
Published online on: June 7, 2022 https://doi.org/10.3892/mmr.2022.12766
Article Number: 250
Copyright: © Lu et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

Metrics: Total Views: 0 (Spandidos Publications: | PMC Statistics: )

Metrics: Total PDF Downloads: 0 (Spandidos Publications: | PMC Statistics: )

Cited By (CrossRef): 0 citations Loading Articles...

Abstract

Macrosomia is a common perinatal complication, with a series of adverse effects on newborns and pregnant women. However, the effects of long non‑coding RNAs (lncRNAs) on non‑diabetic fetal macrosomia (NDFMS) remain unclear. The aim of the present study was to investigate whether aberrant lncRNA expression in the placenta is involved in the pathogenesis of NDFMS and to elucidate its biological mechanisms. The expression profile of lncRNAs in the placentas of pregnant women with NDFMS was investigated using an Agilent Human LncRNA Microarray. Differentially expressed lncRNAs were selected for validation using reverse transcription‑quantitative polymerase chain reaction (RT‑qPCR). Additionally, the function of lncRNA ubiquitin‑specific peptidase 2 antisense RNA 1 (USP2‑AS1) was investigated using a trophoblast cell line. The results revealed that 763 lncRNAs were upregulated and 129 lncRNAs were downregulated in the placentas of women in the NDFMS group (|FC| ≥2.0). A total of 10 lncRNAs (|FC| ≥4.0, signal value ≥50) were selected for validation using two‑stage RT‑qPCR, indicating that the expression trends of the 10 differentially expressed lncRNAs in the NDFMS group (n=8 vs. 8 and 48 vs. 48) were consistent with the microarray data. In addition, a significant downregulation in the levels of lncRNA USP2‑AS1 was observed in both the microarray data and second‑stage verification. The overexpression of lncRNA USP2‑AS1 induced G1 phase cell cycle arrest and the number of cells entering S phase was reduced. In addition, the viability of HTR‑8/SVneo cells was significantly inhibited when lncRNA USP2‑AS1 was overexpressed. Therefore, these findings demonstrated that lncRNAs were significantly differentially expressed in the placentas of pregnant women with NDFMS and that the downregulation of lncRNA USP2‑AS1 may be involved in the pathogenesis of NDFMS, by promoting trophoblast cell viability.

View Figures

View References

1	Kc K, Shakya S and Zhang H: Gestational diabetes mellitus and macrosomia: A literature review. Ann Nutr Metab. 66 (Suppl 2):S14–S20. 2015. View Article : Google Scholar
2	Linder N, Lahat Y, Kogan A, Fridman E, Kouadio F, Melamed N, Yogev Y and Klinger G: Macrosomic newborns of non-diabetic mothers: Anthropometric measurements and neonatal complications. Arch Dis Child Fetal Neonatal Ed. 99:F353–F358. 2014. View Article : Google Scholar : PubMed/NCBI
3	Koyanagi A, Zhang J, Dagvadorj A, Hirayama F, Shibuya K, Souza JP and Gülmezoglu AM: Macrosomia in 23 developing countries: An analysis of a multicountry, facility-based, cross-sectional survey. Lancet. 381:476–483. 2013. View Article : Google Scholar
4	Valvi D, Oulhote Y, Weihe P, Dalgård C, Bjerve KS, Steuerwald U and Grandjean P: Gestational diabetes and offspring birth size at elevated environmental pollutant exposures. Environ Int. 107:205–215. 2017. View Article : Google Scholar
5	Jansson T and Powell TL: Role of placental nutrient sensing in developmental programming. Clin Obstet Gynecol. 56:591–601. 2013. View Article : Google Scholar
6	Jansson T and Powell TL: Role of the placenta in fetal programming: Underlying mechanisms and potential interventional approaches. Clin Sci (Lond). 113:1–13. 2007. View Article : Google Scholar : PubMed/NCBI
7	Tian FY, Wang XM, Xie C, Zhao B, Niu Z, Fan L, Hivert MF and Chen WQ: Placental surface area mediates the association between FGFR2 methylation in placenta and full-term low birth weight in girls. Clin Epigenetics. 10:392018. View Article : Google Scholar : PubMed/NCBI
8	Ilekis JV, Tsilou E, Fisher S, Abrahams VM, Soares MJ, Cross JC, Zamudio S, Illsley NP, Myatt L, Colvis C, et al: Placental origins of adverse pregnancy outcomes: Potential molecular targets: An executive workshop summary of the eunice Kennedy Shriver National institute of child health and human development. Am J Obstet Gynecol. 215 (Suppl 1):S1–S46. 2016. View Article : Google Scholar
9	Longtine MS and Nelson DM: Placental dysfunction and fetal programming: The importance of placental size, shape, histopathology, and molecular composition. Semin Reprod Med. 29:187–196. 2011. View Article : Google Scholar : PubMed/NCBI
10	Lunghi L, Ferretti ME, Medici S, Biondi C and Vesce F: Control of human trophoblast function. Reprod Biol Endocrinol. 5:62007. View Article : Google Scholar
11	Ishihara N, Matsuo H, Murakoshi H, Laoag-Fernandez J, Samoto T and Maruo T: Changes in proliferative potential, apoptosis and Bcl-2 protein expression in cytotrophoblasts and syncytiotrophoblast in human placenta over the course of pregnancy. Endocr J. 47:317–327. 2000. View Article : Google Scholar : PubMed/NCBI
12	Guo D, Jiang H, Chen Y, Yang J, Fu Z, Li J, Han X, Wu X, Xia Y, Wang X, et al: Elevated microRNA-141-3p in placenta of non-diabetic macrosomia regulate trophoblast proliferation. EBioMedicine. 38:154–161. 2018. View Article : Google Scholar : PubMed/NCBI
13	Li J, Fu Z, Jiang H, Chen L, Wu X, Ding H, Xia Y, Wang X, Tang Q and Wu W: IGF2-derived miR-483-3p contributes to macrosomia through regulating trophoblast proliferation by targeting RB1CC1. Mol Hum Reprod. 24:444–452. 2018. View Article : Google Scholar : PubMed/NCBI
14	Lai J, Chen B, Zhang G, Li X, Mok H and Liao N: Molecular characterization of breast cancer: A potential novel immune-related lncRNAs signature. J Transl Med. 18:4162020. View Article : Google Scholar : PubMed/NCBI
15	Sun QM, Hu B, Fu PY, Tang WG, Zhang X, Zhan H, Sun C, He YF, Song K, Xiao YS, et al: Long non-coding RNA 00607 as a tumor suppressor by modulating NF-κB p65/p53 signaling axis in hepatocellular carcinoma. Carcinogenesis. 39:1438–1446. 2018. View Article : Google Scholar : PubMed/NCBI
16	Liang H, Yu T, Han Y, Jiang H, Wang C, You T, Zhao X, Shan H, Yang R, Yang L, et al: LncRNA PTAR promotes EMT and invasion-metastasis in serous ovarian cancer by competitively binding miR-101-3p to regulate ZEB1 expression. Mol Cancer. 17:1192018. View Article : Google Scholar : PubMed/NCBI
17	Knauss JL, Miao N, Kim SN, Nie Y, Shi Y, Wu T, Pinto HB, Donohoe ME and Sun T: Long noncoding RNA Sox2ot and transcription factor YY1 co-regulate the differentiation of cortical neural progenitors by repressing Sox2. Cell Death Dis. 9:7992018. View Article : Google Scholar : PubMed/NCBI
18	Jiang S, Chen Q, Liu H, Gao Y and Yang X, Ren Z, Gao Y, Xiao L, Zhong M, Yu Y and Yang X: Preeclampsia-Associated lncRNA INHBA-AS1 regulates the proliferation, invasion, and migration of placental trophoblast cells. Mol Ther Nucleic Acids. 22:684–695. 2020. View Article : Google Scholar : PubMed/NCBI
19	Chen Q, Jiang S, Liu H, Gao Y, Yang X, Ren Z, Gao Y, Xiao L, Hu H, Yu Y, et al: Association of lncRNA SH3PXD2A-AS1 with preeclampsia and its function in invasion and migration of placental trophoblast cells. Cell Death Dis. 11:5832020. View Article : Google Scholar : PubMed/NCBI
20	Ma Y, Liang X, Wu H, Zhang C and Ma Y: Long noncoding RNA NR_002794 is upregulated in pre-eclampsia and regulates the proliferation, apoptosis and invasion of trophoblast cells. Mol Med Rep. 20:4567–4575. 2019.PubMed/NCBI
21	Pengjie Z, Xionghui C, Yueming Z, Ting X, Na L, Jianying T and Zhice X: LncRNA uc003fir promotes CCL5 expression and negatively affects proliferation and migration of trophoblast cells in preeclampsia. Pregnancy Hypertens. 14:90–96. 2018. View Article : Google Scholar : PubMed/NCBI
22	Zhang Y, Jin F, Li XC, Shen FJ, Ma XL, Wu F, Zhang SM, Zeng WH, Liu XR, Fan JX, et al: The YY1-HOTAIR-MMP2 signaling axis controls trophoblast invasion at the maternal-fetal interface. Mol Ther. 25:2394–2403. 2017. View Article : Google Scholar : PubMed/NCBI
23	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
24	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
25	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
26	Dhanoa JK, Sethi RS, Verma R, Arora JS and Mukhopadhyay CS: Long non-coding RNA: Its evolutionary relics and biological implications in mammals: A review. J Anim Sci Technol. 60:252018. View Article : Google Scholar
27	Du MC, Ouyang YQ, Nie XF, Huang Y and Redding SR: Effects of physical exercise during pregnancy on maternal and infant outcomes in overweight and obese pregnant women: A meta-analysis. Birth. 46:211–221. 2019. View Article : Google Scholar : PubMed/NCBI
28	Poblete JA and Olmos P: Obesity and Gestational diabetes in pregnant care and clinical practice. Curr Vasc Pharmacol. 19:154–164. 2021. View Article : Google Scholar
29	Yang W, Han F, Gao X, Chen Y, Ji L and Cai X: Relationship between gestational weight gain and pregnancy complications or delivery outcome. Sci Rep. 7:125312017. View Article : Google Scholar : PubMed/NCBI
30	Lucarini N, Bottini FG, Borgiani P, Amante A, Gerlini G and Bottini E: Genetic and non genetic factors in the outcome of diabetic pregnancy. J Perinat Med. 22:379–385. 1994. View Article : Google Scholar : PubMed/NCBI
31	Wilcox MA, Newton CS and Johnson IR: Paternal influences on birthweight. Acta Obstet Gynecol Scand. 74:15–18. 1995. View Article : Google Scholar : PubMed/NCBI
32	Tanaka K, Matsushima M, Izawa T, Furukawa S, Kobayashi Y and Iwashita M: Influence of maternal obesity on fetal growth at different periods of pregnancies with normal glucose tolerance. J Obstet Gynaecol Res. 44:691–696. 2018. View Article : Google Scholar : PubMed/NCBI
33	Wallace JM, Horgan GW and Bhattacharya S: Placental weight and efficiency in relation to maternal body mass index and the risk of pregnancy complications in women delivering singleton babies. Placenta. 33:611–618. 2012. View Article : Google Scholar
34	Burton GJ and Jauniaux E: What is the placenta? Am J Obstet Gynecol. 213 (Suppl 4):S6 e1S6-82015. View Article : Google Scholar
35	Gude NM, Roberts CT, Kalionis B and King RG: Growth and function of the normal human placenta. Thromb Res. 114:397–407. 2004. View Article : Google Scholar : PubMed/NCBI
36	Burton GJ and Fowden AL: The placenta: A multifaceted, transient organ. Philos Trans R Soc Lond B Biol Sci. 370:201400662015. View Article : Google Scholar
37	Li T, Hu D and Gong Y: Identification of potential lncRNAs and co-expressed mRNAs in gestational diabetes mellitus by RNA sequencing. J Matern Fetal Neonatal Med. Feb 22–2021.(Epub ahead of print). View Article : Google Scholar
38	Li D, Bao J, Yao J and Li J: lncRNA USP2-AS1 promotes colon cancer progression by modulating Hippo/YAP1 signaling. Am J Transl Res. 12:5670–5682. 2020.PubMed/NCBI
39	Guo B, Yu L, Sun Y, Yao N and Ma L: Long Non-Coding RNA USP2-AS1 accelerates cell proliferation and migration in ovarian cancer by sponging miR-520d-3p and Up-Regulating KIAA1522. Cancer Manag Res. 12:10541–10550. 2020. View Article : Google Scholar : PubMed/NCBI