Expression of mmu-miR-96 in the endometrium during early pregnancy and its regulatory effects on stromal cell apoptosis via Bcl2

Yang,Yuan; Xie,Yi; Wu,Mengyun; Geng,Yanqing; Li,Rong; Xu,Lei; Liu,Xueqing; Pan,Yongquan

doi:10.3892/mmr.2017.6212

Expression of mmu-miR-96 in the endometrium during early pregnancy and its regulatory effects on stromal cell apoptosis via Bcl2

Authors:
- Yuan Yang
- Yi Xie
- Mengyun Wu
- Yanqing Geng
- Rong Li
- Lei Xu
- Xueqing Liu
- Yongquan Pan
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Affiliations: Laboratory of Reproductive Biology, School of Public Health, Chongqing Medical University, Chongqing 400016, P.R. China, Department of Obstetrics and Gynecology, The First Affiliated Hospital of Chongqing Medical University, Chongqing 400016, P.R. China, Laboratory Animal Center, Chongqing Medical University, Chongqing 400016, P.R. China
Published online on: February 17, 2017 https://doi.org/10.3892/mmr.2017.6212
Pages: 1547-1554
Copyright: © Yang et al. This is an open access article distributed under the terms of Creative Commons Attribution License.

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Abstract

Decidualization of endometrial stromal cells is an important feature of implantation and pregnancy. The molecular mechanism underlying decidualization remains unclear, particularly regarding the microRNA (miRNA/miR) regulation of this process. The present study revealed the temporal and spatial distribution of mmu‑miR‑96 in the mouse uterus during early pregnancy by reverse transcription‑quantitative polymerase chain reaction and in situ hybridization. In addition, primary stromal cells were isolated from the mouse uterus and used to explore the role of mmu‑miR‑96 in decidualization. The results demonstrated that mmu‑miR‑96 was highly expressed in stromal cells during pregnancy, and was upregulated at implantation sites. In addition, mmu‑miR‑96 was strongly expressed during decidualization, which indicates that it may serve a role in the decidualization of stromal cells. Based on existing reports, mmu‑miR‑96 participates in apoptosis; therefore the present study investigated its effects on the apoptosis of primary endometrial stromal cells. The results indicated that overexpression of mmu‑miR‑96 may induce apoptosis of stromal cells. In further studies regarding the underlying mechanism, the target genes of mmu‑miR‑96 were screened by bioinformatics analysis, and it was confirmed that B‑cell lymphoma 2, an anti‑apoptotic gene, was the target of mmu‑miR‑96, as determined using a reporter gene assay. In conclusion, the present study suggested that mmu‑miR‑96 participates in the decidualization of endometrial stromal cells in mice, thereby serving a key role in pregnancy.

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1	Cha J, Sun X and Dey SK: Mechanisms of implantation: Strategies for successful pregnancy. Nat Med. 18:1754–1767. 2012. View Article : Google Scholar : PubMed/NCBI
2	Wang H and Dey SK: Roadmap to embryo implantation: Clues from mouse models. Nat Rev Genet. 7:185–199. 2006. View Article : Google Scholar : PubMed/NCBI
3	Parkes AS: The Functions of the Corpus Luteum. II.-The Experimental Production of Placentomata in the Mouse. Proceedings of the Royal Society of London Series B Containing Papers of a Biological Character. 104:183–188. 1929. View Article : Google Scholar
4	Shimizu A, Maruyama T, Tamaki K, Uchida H, Asada H and Yoshimura Y: Impairment of decidualization in SRC-deficient mice. Biol Reprod. 73:1219–1227. 2005. View Article : Google Scholar : PubMed/NCBI
5	Eyal O, Jomain JB, Kessler C, Goffin V and Handwerger S: Autocrine prolactin inhibits human uterine decidualization: A novel role for prolactin. Biol Reprod. 76:777–783. 2007. View Article : Google Scholar : PubMed/NCBI
6	Chakrabarty A, Tranguch S, Daikoku T, Jensen K, Furneaux H and Dey SK: MicroRNA regulation of cyclooxygenase-2 during embryo implantation. Proc Natl Acad Sci USA. 104:15144–15149. 2007. View Article : Google Scholar : PubMed/NCBI
7	Hu SJ, Ren G, Liu JL, Zhao ZA, Yu YS, Su RW, Ma XH, Ni H, Lei W and Yang ZM: MicroRNA expression and regulation in mouse uterus during embryo implantation. J Biol Chem. 283:23473–23484. 2008. View Article : Google Scholar : PubMed/NCBI
8	Shen LJ, He JL, Yang DH, Ding YB, Chen XM, Geng YQ, Liu SJ, Liu XQ and Wang YX: Mmu-microRNA-200a overexpression leads to implantation defect by targeting phosphatase and tensin homolog in mouse uterus. Reprod Sci. 20:1518–1528. 2013. View Article : Google Scholar : PubMed/NCBI
9	Singh SK, Pal Bhadra M, Girschick HJ and Bhadra U: MicroRNAs-micro in size but macro in function. FEBS J. 275:4929–4944. 2008. View Article : Google Scholar : PubMed/NCBI
10	Zhao Y and Srivastava D: A developmental view of microRNA function. Trends Biochem Sci. 32:189–197. 2007. View Article : Google Scholar : PubMed/NCBI
11	Chen K, Chen X, He J, Ding Y, Geng Y, Liu S, Liu X and Wang Y: Mouse endometrium temporal and spatial expression mRNA and microRNA associated with embryo implantation. Reprod Sci. 22:1399–1408. 2015. View Article : Google Scholar : PubMed/NCBI
12	Han Y, Chen J, Zhao X, Liang C, Wang Y, Sun L, Jiang Z, Zhang Z, Yang R, Chen J, et al: MicroRNA expression signatures of bladder cancer revealed by deep sequencing. PLoS One. 6:e182862011. View Article : Google Scholar : PubMed/NCBI
13	Yamada Y, Enokida H, Kojima S, Kawakami K, Chiyomaru T, Tatarano S, Yoshino H, Kawahara K, Nishiyama K, Seki N and Nakagawa M: MiR-96 and miR-183 detection in urine serve as potential tumor markers of urothelial carcinoma: Correlation with stage and grade, and comparison with urinary cytology. Cancer Sci. 102:522–529. 2011. View Article : Google Scholar : PubMed/NCBI
14	Nymark P, Guled M, Borze I, Faisal A, Lahti L, Salmenkivi K, Kettunen E, Anttila S and Knuutila S: Integrative analysis of microRNA, mRNA and aCGH data reveals asbestos- and histology-related changes in lung cancer. Genes Chromosomes Cancer. 50:585–597. 2011. View Article : Google Scholar : PubMed/NCBI
15	Lin H, Dai T, Xiong H, Zhao X, Chen X, Yu C, Li J, Wang X and Song L: Unregulated miR-96 induces cell proliferation in human breast cancer by downregulating transcriptional factor FOXO3a. PLoS One. 5:e157972010. View Article : Google Scholar : PubMed/NCBI
16	Myatt SS, Wang J, Monteiro LJ, Christian M, Ho KK, Fusi L, Dina RE, Brosens JJ, Ghaem-Maghami S and Lam EW: Definition of microRNAs that repress expression of the tumor suppressor gene FOXO1 in endometrial cancer. Cancer Res. 70:367–377. 2010. View Article : Google Scholar : PubMed/NCBI
17	Pineau P, Volinia S, McJunkin K, Marchio A, Battiston C, Terris B, Mazzaferro V, Lowe SW, Croce CM and Dejean A: miR-221 overexpression contributes to liver tumorigenesis. Proc Natl Acad Sci USA. 107:264–269. 2010. View Article : Google Scholar : PubMed/NCBI
18	Guo Y, Liu H, Zhang H, Shang C and Song Y: miR-96 regulates FOXO1-mediated cell apoptosis in bladder cancer. Oncol Lett. 4:561–565. 2012.PubMed/NCBI
19	Guttilla IK and White BA: Coordinate regulation of FOXO1 by miR-27a, miR-96, and miR-182 in breast cancer cells. J Biol Chem. 284:23204–23216. 2009. View Article : Google Scholar : PubMed/NCBI
20	Li J, Li P, Chen T, Gao G, Chen X, Du Y, Zhang R, Yang R, Zhao W, Dun S, et al: Expression of microRNA-96 and its potential functions by targeting FOXO3 in non-small cell lung cancer. Tumour Biol. 36:685–692. 2014. View Article : Google Scholar : PubMed/NCBI
21	Ma XH, Hu SJ, Ni H, Zhao YC, Tian Z, Liu JL, Ren G, Liang XH, Yu H, Wan P and Yang ZM: Serial analysis of gene expression in mouse uterus at the implantation site. J Biol Chem. 281:9351–9360. 2006. View Article : Google Scholar : PubMed/NCBI
22	Tan Y, Li M, Cox S, Davis MK, Tawfik O, Paria BC and Das SK: HB-EGF directs stromal cell polyploidy and decidualization via cyclin D3 during implantation. Dev Biol. 265:181–195. 2004. View Article : Google Scholar : PubMed/NCBI
23	Liao XG, Li YL, Gao RF, Geng YQ, Chen XM, Liu XQ, Ding YB, Mu XY, Wang YX and He JL: Folate deficiency decreases apoptosis of endometrium decidual cells in pregnant mice via the mitochondrial pathway. Nutrients. 7:1916–1932. 2015. View Article : Google Scholar : PubMed/NCBI
24	Li Q, Kannan A, Wang W, Demayo FJ, Taylor RN, Bagchi MK and Bagchi IC: Bone morphogenetic protein 2 functions via a conserved signaling pathway involving Wnt4 to regulate uterine decidualization in the mouse and the human. J Biol Chem. 282:31725–31732. 2007. View Article : Google Scholar : PubMed/NCBI
25	Liang XH, Zhao ZA, Deng WB, Tian Z, Lei W, Xu X, Zhang XH, Su RW and Yang ZM: Estrogen regulates amiloride-binding protein 1 through CCAAT/enhancer-binding protein-beta in mouse uterus during embryo implantation and decidualization. Endocrinology. 151:5007–5016. 2010. View Article : Google Scholar : PubMed/NCBI
26	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
27	Wu R, Tang S, Wang M, Xu X, Yao C and Wang S: MicroRNA-497 induces apoptosis and suppresses proliferation via the Bcl-2/Bax-Caspase9-caspase3 pathway and cyclin D2 protein in HUVECs. PLoS One. 11:e01670522016. View Article : Google Scholar : PubMed/NCBI
28	Liu Z, Jiang R, Yuan S, Wang N, Feng Y, Hu G, Zhu X, Huang K, Ma J, Xu G, et al: Integrated analysis of DNA methylation and RNA transcriptome during in vitro differentiation of human pluripotent stem cells into retinal pigment epithelial cells. PLoS One. 9:e914162014. View Article : Google Scholar : PubMed/NCBI
29	Nashta AA Fouladi, Andreu CV, Nijjar N, Heath JK and Kimber SJ: Role of leukemia inhibitor factor (LIF) in decidualisation of murine uterine stromal cells in vitro. J Endocrinol. 181:477–492. 2004. View Article : Google Scholar : PubMed/NCBI
30	Evans MJ and Kaufman MH: Establishment in culture of pluripotential cells from mouse embryos. Nature. 292:154–156. 1981. View Article : Google Scholar : PubMed/NCBI
31	Estella C, Herrer I, Moreno-Moya JM, Quiñonero A, Martínez S, Pellicer A and Simón C: miRNA signature and Dicer requirement during human endometrial stromal decidualization in vitro. PLoS One. 7:e410802012. View Article : Google Scholar : PubMed/NCBI
32	Zhang Q, Zhang H, Jiang Y, Xue B, Diao Z, Ding L, Zhen X, Sun H, Yan G and Hu Y: MicroRNA-181a is involved in the regulation of human endometrial stromal cell decidualization by inhibiting Kruppel-like factor 12. Reprod Biol Endocrinol. 13:232015. View Article : Google Scholar : PubMed/NCBI
33	Boeddeker SJ and Hess AP: The role of apoptosis in human embryo implantation. J Reprod Immunol. 108:114–122. 2015. View Article : Google Scholar : PubMed/NCBI
34	Correia-da-Silva G, Bell SC, Pringle JH and Teixeira NA: Patterns of expression of Bax, Bcl-2 and Bcl-x(L) in the implantation site in rat during pregnancy. Placenta. 26:796–806. 2005. View Article : Google Scholar : PubMed/NCBI
35	Joswig A, Gabriel HD, Kibschull M and Winterhager E: Apoptosis in uterine epithelium and decidua in response to implantation: Evidence for two different pathways. Reprod Biol Endocrinol. 1:442003. View Article : Google Scholar : PubMed/NCBI