Top

Reproductive Biology and Endocrinology

Published in:

Open Access 01-12-2015 | Research

Uterine NDRG2 expression is increased at implantation sites during early pregnancy in mice, and its down-regulation inhibits decidualization of mouse endometrial stromal cells

Authors: Yan Gu, Xuan Zhang, Qian Yang, Jian-mei Wang, Ya-ping He, Zhao-gui Sun, Hui-qin Zhang, Jian Wang

Published in: Reproductive Biology and Endocrinology | Issue 1/2015

Abstract

Background

N-myc down-regulated gene 2 (NDRG2) is a tumor suppressor involved in cell proliferation and differentiation. The aim of this study was to determine the uterine expression pattern of this gene during early pregnancy in mice.

Methods

Uterine NDRG2 mRNA and protein expression levels were determined by RT-PCR and Western blot analyses, respectively, during the peri-implantation period in mice. Immunohistochemical (IHC) analysis was performed to examine the spatial localization of NDRG2 expression in mouse uterine tissues. The in vitro decidualization model of mouse endometrial stromal cells (ESCs) was used to evaluate decidualization of ESCs following NDRG2 knock down by small interfering RNA (siRNA). Statistical significance was analyzed by one-way ANOVA using SPSS 19.0 software.

Results

Uterine NDRG2 gene expression was significantly up-regulated and was predominantly localized to the secondary decidual zone on days 5 and 8 of pregnancy in mice. Its increased expression was associated with artificial decidualization as well as the activation of delayed implantation. Furthermore, uterine NDRG2 expression was induced by estrogen and progesterone treatments. The in vitro decidualization of mouse ESCs was accompanied by up-regulation of NDRG2 expression, and knock down of its expression in these cells by siRNA inhibited the decidualization process.

Conclusions

These results suggest that NDRG2 might play an important role in the process of decidualization during early pregnancy.

Aplin JD. Embryo implantation: the molecular mechanism remains elusive. Reprod Biomed Online. 2006;13(6):833–9.CrossRefPubMed

Harun R, Ruban L, Matin M, Draper J, Jenkins NM, Liew GC, et al. Cytotrophoblast stem cell lines derived from human embryonic stem cells and their capacity to mimic invasive implantation events. Hum Reprod. 2006;21(6):1349–58. doi:10.1093/humrep/del017.CrossRefPubMed

Ferretti C, Bruni L, Dangles-Marie V, Pecking AP, Bellet D. Molecular circuits shared by placental and cancer cells, and their implications in the proliferative, invasive and migratory capacities of trophoblasts. Hum Reprod Update. 2007;13(2):121–41. doi:10.1093/humupd/dml048.CrossRefPubMed

Cameo P, Srisuparp S, Strakova Z, Fazleabas AT. Chorionic gonadotropin and uterine dialogue in the primate. Reprod Biol Endocrinol. 2004;2:50. doi:10.1186/1477-7827-2-50.PubMedCentralCrossRefPubMed

Loke YW, King A, Burrows TD. Decidua in human implantation. Hum Reprod. 1995;10 Suppl 2:14–21.CrossRefPubMed

Gellersen B, Brosens IA, Brosens JJ. Decidualization of the human endometrium: mechanisms, functions, and clinical perspectives. Semin Reprod Med. 2007;25(6):445–53. doi:10.1055/s-2007-991042.CrossRefPubMed

Ma M, Zhao K, Wu W, Sun R, Fei J. Dynamic expression of N-myc in mouse embryonic development using an enhanced green fluorescent protein reporter gene in the N-myc locus. Dev Growth Differ. 2014;56(2):152–60. doi:10.1111/dgd.12115.CrossRefPubMed

Yao L, Zhang J, Liu X. NDRG2: a Myc-repressed gene involved in cancer and cell stress. Acta Biochim Biophys Sin. 2008;40(7):625–35.CrossRefPubMed

Liu N, Wang L, Liu X, Yang Q, Zhang J, Zhang W, et al. Promoter methylation, mutation, and genomic deletion are involved in the decreased NDRG2 expression levels in several cancer cell lines. Biochem Biophys Res Commun. 2007;358(1):164–9. doi:10.1016/j.bbrc.2007.04.089.CrossRefPubMed

10.

Hu XL, Liu XP, Lin SX, Deng YC, Liu N, Li X, et al. NDRG2 expression and mutation in human liver and pancreatic cancers. World J Gastroenterol. 2004;10(23):3518–21.PubMed

11.

Assamaki R, Sarlomo-Rikala M, Lopez-Guerrero JA, Lasota J, Andersson LC, Llombart-Bosch A, et al. Array comparative genomic hybridization analysis of chromosomal imbalances and their target genes in gastrointestinal stromal tumors. Gene Chromosome Canc. 2007;46(6):564–76. doi:10.1002/gcc.20439.CrossRef

12.

Hu XL, Liu XP, Deng YC, Lin SX, Wu L, Zhang J, et al. Expression analysis of the NDRG2 gene in mouse embryonic and adult tissues. Cell Tissue Res. 2006;325(1):67–76. doi:10.1007/s00441-005-0137-5.CrossRefPubMed

13.

Burchfield JG, Lennard AJ, Narasimhan S, Hughes WE, Wasinger VC, Corthals GL, et al. Akt mediates insulin-stimulated phosphorylation of Ndrg2: evidence for cross-talk with protein kinase C theta. J Biol Chem. 2004;279(18):18623–32. doi:10.1074/jbc.M401504200.CrossRefPubMed

14.

Murray JT, Campbell DG, Morrice N, Auld GC, Shpiro N, Marquez R, et al. Exploitation of KESTREL to identify NDRG family members as physiological substrates for SGK1 and GSK3. Biochem J. 2004;384(Pt 3):477–88. doi:10.1042/BJ20041057.PubMedCentralCrossRefPubMed

15.

Lyu SY, Choi JH, Lee HJ, Park WB, Kim GJ. Korean mistletoe lectin promotes proliferation and invasion of trophoblast cells through regulation of Akt signaling. Reprod Toxicol. 2013;39:33–9. doi:10.1016/j.reprotox.2013.03.011.CrossRefPubMed

16.

Gouon-Evans V, Pollard JW. Eotaxin is required for eosinophil homing into the stroma of the pubertal and cycling uterus. Endocrinology. 2001;142(10):4515–21. doi:10.1210/endo.142.10.8459.CrossRefPubMed

17.

O'Sullivan CM, Liu SY, Rancourt SL, Rancourt DE. Regulation of the strypsin-related proteinase ISP2 by progesterone in endometrial gland epithelium during implantation in mice. Reproduction. 2001;122(2):235–44.CrossRefPubMed

18.

Hayashi K, Erikson DW, Tilford SA, Bany BM, Maclean 2nd JA, Rucker 3rd EB, et al. Wnt genes in the mouse uterus: potential regulation of implantation. Biol Reprod. 2009;80(5):989–1000. doi:10.1095/biolreprod.108.075416.PubMedCentralCrossRefPubMed

19.

Chen Y, Ni H, Ma XH, Hu SJ, Luan LM, Ren G, et al. Global analysis of differential luminal epithelial gene expression at mouse implantation sites. J Mol Endocrinol. 2006;37(1):147–61. doi:10.1677/jme.1.02009.CrossRefPubMed

20.

Maurya VK, Jha RK, Kumar V, Joshi A, Chadchan S, Mohan JJ, et al. Transforming growth factor-beta 1 (TGF-B1) liberation from its latent complex during embryo implantation and its regulation by estradiol in mouse. Biol Reprod. 2013;89(4):84. doi:10.1095/biolreprod.112.106542.CrossRefPubMed

21.

Hu SJ, Ren G, Liu JL, Zhao ZA, Yu YS, Su RW, et al. MicroRNA expression and regulation in mouse uterus during embryo implantation. J Biol Chem. 2008;283(34):23473–84. doi:10.1074/jbc.M800406200.CrossRefPubMed

22.

Nautiyal J, Kumar PG, Laloraya M. 17Beta-estradiol induces nuclear translocation of CrkL at the window of embryo implantation. Biochem Biophys Res Commun. 2004;318(1):103–12. doi:10.1016/j.bbrc.2004.04.005.CrossRefPubMed

23.

Hamatani T, Daikoku T, Wang H, Matsumoto H, Carter MG, Ko MS, et al. Global gene expression analysis identifies molecular pathways distinguishing blastocyst dormancy and activation. Proc Natl Acad Sci U S A. 2004;101(28):10326–31. doi:10.1073/pnas.0402597101.PubMedCentralCrossRefPubMed

24.

Smith SE, French MM, Julian J, Paria BC, Dey SK, Carson DD. Expression of heparan sulfate proteoglycan (perlecan) in the mouse blastocyst is regulated during normal and delayed implantation. Dev Biol. 1997;184(1):38–47. doi:10.1006/dbio.1997.8521.CrossRefPubMed

25.

Li Q, Kannan A, Wang W, Demayo FJ, Taylor RN, Bagchi MK, et al. 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. 2007;282(43):31725–32. doi:10.1074/jbc.M704723200.CrossRefPubMed

26.

Kimura F, Takakura K, Takebayashi K, Ishikawa H, Kasahara K, Goto S, et al. Messenger ribonucleic acid for the mouse decidual prolactin is present and induced during in vitro decidualization of endometrial stromal cells. Gynecol Endocrinol. 2001;15(6):426–32.CrossRefPubMed

27.

Singh M, Chaudhry P, Asselin E. Bridging endometrial receptivity and implantation: network of hormones, cytokines, and growth factors. J Endocrinol. 2011;210(1):5–14. doi:10.1530/JOE-10-0461.CrossRefPubMed

28.

Salgado RM, Favaro RR, Zorn TM. Modulation of small leucine-rich proteoglycans (SLRPs) expression in the mouse uterus by estradiol and progesterone. Reprod Biol Endocrinol. 2011;9:22. doi:10.1186/1477-7827-9-22.PubMedCentralCrossRefPubMed

29.

Li Y, Yang J, Li S, Zhang J, Zheng J, Hou W, et al. N-myc downstream-regulated gene 2, a novel estrogen-targeted gene, is involved in the regulation of Na+/K + -ATPase. J Biol Chem. 2011;286(37):32289–99. doi:10.1074/jbc.M111.247825.PubMedCentralCrossRefPubMed

30.

Ma YL, Qin P, Feng DY, Li Y, Zhang LX, Liu ZY, et al. Estrogen regulates the expression of Ndrg2 in astrocytes. Brain research. 2014;1569:1–8. doi:10.1016/j.brainres.2014.04.036.CrossRefPubMed

31.

Horvat B. Cyclic changes in the expression of endometrial proteins of the mouse during the normal estrus cycle. In vivo. 1991;5(4):345–7.PubMed

32.

Das SK. Cell cycle regulatory control for uterine stromal cell decidualization in implantation. Reproduction. 2009;137(6):889–99. doi:10.1530/REP-08-0539.CrossRefPubMed

33.

Paulson RJ. Hormonal induction of endometrial receptivity. Fertil Steril. 2011;96(3):530–5. doi:10.1016/j.fertnstert.2011.07.1097.CrossRefPubMed

34.

King AE, Critchley HO. Oestrogen and progesterone regulation of inflammatory processes in the human endometrium. J Steroid Biochem Mol Biol. 2010;120(2-3):116–26. doi:10.1016/j.jsbmb.2010.01.003.CrossRefPubMed

35.

Ramathal CY, Bagchi IC, Taylor RN, Bagchi MK. Endometrial decidualization: of mice and men. Semin Reprod Med. 2010;28(1):17–26. doi:10.1055/s-0029-1242989.PubMedCentralCrossRefPubMed

36.

Erlebacher A. Immunology of the maternal-fetal interface. Annu Rev Immunol. 2013;31:387–411. doi:10.1146/annurev-immunol-032712-100003.CrossRefPubMed

37.

Choi SC, Kim KD, Kim JT, Kim JW, Lee HG, Kim JM, et al. Expression of human NDRG2 by myeloid dendritic cells inhibits down-regulation of activated leukocyte cell adhesion molecule (ALCAM) and contributes to maintenance of T cell stimulatory activity. J Leukoc Biol. 2008;83(1):89–98. doi:10.1189/jlb.0507300.CrossRefPubMed

38.

Mori M, Kitazume M, Ose R, Kurokawa J, Koga K, Osuga Y, et al. Death effector domain-containing protein (DEDD) is required for uterine decidualization during early pregnancy in mice. J Clin Invest. 2011;121(1):318–27. doi:10.1172/JCI44723.PubMedCentralCrossRefPubMed

39.

Salker MS, Christian M, Steel JH, Nautiyal J, Lavery S, Trew G, et al. Deregulation of the serum- and glucocorticoid-inducible kinase SGK1 in the endometrium causes reproductive failure. Nature medicine. 2011;17(11):1509–13. doi:10.1038/nm.2498.CrossRefPubMed

40.

Oreshkova T, Dimitrov R, Mourdjeva M. A cross-talk of decidual stromal cells, trophoblast, and immune cells: a prerequisite for the success of pregnancy. Am J Reprod Immunol. 2012;68(5):366–73. doi:10.1111/j.1600-0897.2012.01165.x.CrossRefPubMed

41.

Basak S, Dhar R, Das C. Steroids modulate the expression of alpha4 integrin in mouse blastocysts and uterus during implantation. Biol Reprod. 2002;66(6):1784–9.CrossRefPubMed

42.

Wang H, Bocca S, Anderson S, Yu L, Rhavi BS, Horcajadas J, et al. Sex steroids regulate epithelial-stromal cell cross talk and trophoblast attachment invasion in a three-dimensional human endometrial culture system. Tissue Eng Part C Methods. 2013;19(9):676–87. doi:10.1089/ten.TEC.2012.0616.CrossRefPubMed

Title: Uterine NDRG2 expression is increased at implantation sites during early pregnancy in mice, and its down-regulation inhibits decidualization of mouse endometrial stromal cells
Authors: Yan Gu
Xuan Zhang
Qian Yang
Jian-mei Wang
Ya-ping He
Zhao-gui Sun
Hui-qin Zhang
Jian Wang
Publication date: 01-12-2015
Publisher: BioMed Central
Published in: Reproductive Biology and Endocrinology / Issue 1/2015
Electronic ISSN: 1477-7827
DOI: https://doi.org/10.1186/s12958-015-0047-7

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Uterine NDRG2 expression is increased at implantation sites during early pregnancy in mice, and its down-regulation inhibits decidualization of mouse endometrial stromal cells

Abstract

Background

Methods

Results

Conclusions

At a glance: The ONWARDS insulin icodec trials

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

Please log in to get access to this content

Other articles of this Issue 1/2015

Predictive value of the time interval between embryo loading and transfer for IVF/ICSI success: a prospective cohort study

Influence of ejaculation frequency on seminal parameters

Isoform 165 of vascular endothelial growth factor in collagen matrix improves ovine cryopreserved ovarian tissue revascularisation after xenotransplantation in mice

Offspring subcutaneous adipose markers are sensitive to the timing of maternal gestational weight gain

The involvement of osteopontin and matrix metalloproteinase- 9 in the migration of endometrial epithelial cells in patients with endometriosis

Aberrant expression of Notch1/numb/snail signaling, an epithelial mesenchymal transition related pathway, in adenomyosis