Abstract
Decidualization is crucial for successful pregnancy in mice and humans. Although many essential molecular modulators have been identified during decidualization, the precise molecular mechanism of uterine decidualization remains largely unknown. Our previous research indicates that luman recruiting factor (LRF) is strongly expressed in decidual uteri of mice on days 6-8 of pregnancy. In this study, our aim is to determine the biological functions of LRF during decidualization in mice. We used the shLRF lentivirus to attenuate the expression of LRF, which significantly reduced the weight and size of implantation sites on days 7-8 of pregnancy. In a stromal cell culture model, LRF mRNA and protein levels increased significantly during stromal cell decidualization induced by estrogen and progesterone. LRF silencing resulted in the decidual markers decidual prolactin-related protein, insulin-like growth factor-binding protein 1 and progesterone receptor being dramatically reduced, and the decidual process was significantly inhibited. Cell-cycle analysis and cell apoptosis analysis revealed that, although no obvious apoptosis occurred in shLRF-lentivirus-infected stromal cells during decidualization, proliferation was inhibited via S-phase cell-cycle arrest, and the mitotic activity of uterine stromal cells was inhibited. An examination of cell-cycle regulatory factors indicated that the mRNA expression levels of cyclin A and cyclin B1 were significantly down-regulated after treatment with shLRF lentivirus. Thus, LRF seems to be involved in the regulation of decidualization during pregnancy by modulating the expression of the key cell-cycle regulatory factors cyclin A and cyclin B1.
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References
Audas TE, Li Y, Liang G, Lu R (2008) A novel protein, Luman/CREB3 recruitment factor, inhibits Luman activation of the unfolded protein response. Mol Cell Biol 28:3952–3966
Brandeis M, Rosewell I, Carrington M, Crompton T, Jacobs MA, Kirk J, Gannon J, Hunt T (1998) Cyclin B2-null mice develop normally and are fertile whereas cyclin B1-null mice die in utero. Proc Natl Acad Sci U S A 95:4344–4349
Chen F, Lin PF, Li X, Sun J, Zhang Z, Du E, Wang A, Jin YP (2014) Construction and expression of lentiviral vectors encoding recombinant mouse CREBZF in NIH 3T3 cells. Plasmid 76C:24–31
Dey S, Lim H, Das SK, Reese J, Paria B, Daikoku T, Wang H (2004) Molecular cues to implantation. Endocr Rev 25:341–373
Filant J, Spencer TE (2014) Uterine glands: biological roles in conceptus implantation, uterine receptivity and decidualization. Int J Dev Biol 58:107–116
Gu Y, Gibori G (1995) Isolation, culture, and characterization of the two cell subpopulations forming the rat decidua: differential gene expression for activin, follistatin, and decidual prolactin-related protein. Endocrinology 136:2451–2458
Hustin J, Philippe E, Teisner B, Grudzinskas J (1994) Immunohistochemical localization of two endometrial proteins in the early days of human pregnancy. Placenta 15:701–708
Kajihara T, Brosens JJ, Ishihara O (2013) The role of FOXO1 in the decidual transformation of the endometrium and early pregnancy. Med Mol Morphol 46:61–68
Kommagani R, Szwarc MM, Kovanci E, Creighton CJ, O'Malley BW, Demayo FJ, Lydon JP (2014) A murine uterine transcriptome, responsive to steroid receptor coactivator-2, reveals transcription factor 23 as essential for decidualization of human endometrial stromal cells. Biol Reprod 90:75
Kover K, Liang L, Andrews GK, Dey SK (1995) Differential expression and regulation of cytokine genes in the mouse uterus. Endocrinology 136:1666–1673
Lan X, Jin Y, Yang Y, Lin P, Hu L, Cui C, Li Q, Li X, Wang A (2013) Expression and localization of Luman RNA and protein during mouse implantation and decidualization. Theriogenology 80:e131–e132
Lin P, Chen F, Wang N, Wang X, Li X, Zhou J, Jin Y, Wang A (2013) CREBZF expression and hormonal regulation in the mouse uterus. Reprod Biol Endocrinol 11:110
Lin PF, Jin YP, Lan XL, Yang YZ, Chen FL, Wang N, Li X, Sun YJ, Wang AH (2014) GRP78 expression and regulation in the mouse uterus during embryo implantation. J Mol Histol 45:259–268
Lu R, Yang P, O'Hare P, Misra V (1997) Luman, a new member of the CREB/ATF family, binds to Herpes simplex virus VP16-associated host cellular factor. Mol Cell Biol 17:5117–5126
Martyn AC, Choleris E, Gillis DJ, Armstrong JN, Amor TR, McCluggage AR, Turner PV, Liang G, Cai K, Lu R (2012) Luman/CREB3 recruitment factor regulates glucocorticoid receptor activity and is essential for prolactin-mediated maternal instinct. Mol Cell Biol 32:5140–5150
Morgan DO (1997) Cyclin-dependent kinases: engines, clocks, and microprocessors. Annu Rev Cell Dev Biol 13:261–291
Mori M, Kitazume M, Ose R, Kurokawa J, Koga K, Osuga Y, Arai S, Miyazaki T (2011) Death effector domain-containing protein (DEDD) is required for uterine decidualization during early pregnancy in mice. J Clin Invest 121:318–327
Muter J, Lucas ES, Chan YW, Brighton PJ, Moore JD, Lacey L, Quenby S, Lam EW, Brosens JJ (2015) The clock protein period 2 synchronizes mitotic expansion and decidual transformation of human endometrial stromal cells. FASEB J 29:1603–1614
Orwig KE, Ishimura R, Muller H, Liu B, Soares MJ (1997) Identification and characterization of a mouse homolog for decidual/trophoblast prolactin-related protein. Endocrinology 138:5511–5517
Richardson H, Lew DJ, Henze M, Sugimoto K, Reed SI (1992) Cyclin-B homologs in Saccharomyces cerevisiae function in S phase and in G2. Genes Dev 6:2021–2034
Sroga JM, Ma X, Das SK (2012) Developmental regulation of decidual cell polyploidy at the site of implantation. Front Biosci (Schol Ed) 4:1475–1486
Su RW, Strug MR, Joshi NR, Jeong JW, Miele L, Lessey BA, Young SL, Fazleabas AT (2015) Decreased notch pathway signaling in the endometrium of women with endometriosis impairs decidualization. J Clin Endocrinol Metab 100:E433–E442
Tan J, Raja S, Davis MK, Tawfik O, Dey SK, Das SK (2002) Evidence for coordinated interaction of cyclin D3 with p21 and cdk6 in directing the development of uterine stromal cell decidualization and polyploidy during implantation. Mech Dev 111:99–113
Tan Y, Li M, Cox S, Davis MK, Tawfik O, Paria BC, Das SK (2004) HB-EGF directs stromal cell polyploidy and decidualization via cyclin D3 during implantation. Dev Biol 265:181–195
Tang L, Wang TT, Wu YT, Zhou CY, Huang HF (2009) High expression levels of cyclin B1 and Polo-like kinase 1 in ectopic endometrial cells associated with abnormal cell cycle regulation of endometriosis. Fertil Steril 91:979–987
Tong XM, Lin XN, Song T, Liu L, Zhang SY (2010) Calcium-binding protein S100P is highly expressed during the implantation window in human endometrium. Fertil Steril 94:1510–1518
Wang Q, Lu J, Zhang S, Wang S, Wang W, Wang B, Wang F, Chen Q, Duan E, Leitges M, Kispert A, Wang H (2012) Wnt6 is essential for stromal cell proliferation during decidualization in mice. Biol Reprod 88:5–5
Wang W, Li QX, Bagchi IC, Bagchi MK (2010) The CCAAT/enhancer binding protein beta is a critical regulator of steroid-induced mitotic expansion of uterine stromal cells during decidualization. Endocrinology 151:3929–3940
Yang Y, Jin Y, Lin P, Hu L, Cui C, Li X, Li Q, Wang A (2013a) The expression and localization of LRF in the female reproductive tract of cycling mice throughout the estrous cycle. J Immunoassay Immunochem 34:313–322
Yang YZ, Jin YP, Martyn AC, Lin PF, Song YJ, Chen FL, Hu LY, Cui CC, Li X, Li Q, Lu R, Wang AH (2013b) Expression pattern implicates a potential role for luman recruitment factor in the process of implantation in uteri and development of preimplantation embryos in mice. J Reprod Dev 59:245–251
Yang YZ, Lin PF, Chen FL, Wang AH, Lan LL, Song YJ, Jin YP (2013c) Luman recruiting factor regulates endoplasmic reticulum stress in mouse ovarian granulosa cell apoptosis. Theriogenology 79:633–639
Zhang HL, Chen XM, Zhao Y, Gao RF, Geng YQ, Ding YB, Liu XQ, Wang YX, He JL (2015) The homologous genes Vangl1 and Vangl2 are required for embryo implantation in the uterus of mice during early pregnancy. Gene 555:140–149
Zhang YY, Chen H, Sun C, Wang HZ, Liu ML, Li YY, Nie XL, Du MR, Li DJ, Zhang JP (2014) Expression and functional characterization of NOD2 in decidual stromal cells isolated during the first trimester of pregnancy. PloS One 9:e99612
Zhu H, Hou CC, Luo LF, Hu YJ, Yang WX (2014) Endometrial stromal cells and decidualized stromal cells: origins, transformation and functions. Gene 551:1–14
Acknowledgments
We thank Dr. Enqi Du for providing technical assistance and the recombinant lentiviral packaging system.
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This study was funded by the National Natural Science Foundation of China (31372499), the Specialized Natural Science Basic Research Plan in Shaanxi Province of China (2012JQ3001) and Fundamental Research Funds for the Central Universities (QN2011105).
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Li, X., Lin, P., Chen, F. et al. Luman recruiting factor is involved in stromal cell proliferation during decidualization in mice. Cell Tissue Res 365, 437–447 (2016). https://doi.org/10.1007/s00441-016-2392-z
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DOI: https://doi.org/10.1007/s00441-016-2392-z