In endometrial epithelial cells, progesterone (P4) functions in regulating the cell structure and opposing the effects of estrogen. However, the mechanisms of P4 that oppose the effects of estrogen remain unclear. MicroRNAs (miRNAs) are important posttranscriptional regulators that are involved in various physiological and pathological processes. Whether P4 directly induces miRNA expression to antagonize estrogen in endometrial epithelium is unclear. In this study, total RNAs were extracted from endometrial epithelium of ovariectomized mice, which were treated with estrogen alone or a combination of estrogen and P4. MicroRNA high-throughput sequencing with bioinformatics analysis was used to identify P4-induced miRNAs, predict their potential target genes, and analyze their possible biological functions. We observed that 146 mature miRNAs in endometrial epithelial cells were significantly upregulated by P4. These miRNAs were extensively involved in multiple biological processes. The miRNA-145a demonstrated a possible function in the antiproliferative action of P4 on endometrial epithelial cells.
TongWPollardJW. Progesterone inhibits estrogen-induced cyclin D1 and cdk4 nuclear translocation, cyclin E- and cyclin A-cdk2 kinase activation, and cell proliferation in uterine epithelial cells in mice. Mol Cell Biol. 1999;19
(3):2251–2264.
2.
GrahamJDClarkeCL. Physiological action of progesterone in target tissues. Endocr Rev. 1997;18
(4):502–519.
3.
ChwaliszKStockemannKFritzemeierKHFuhrmannU. Modulation of oestrogenic effects by progesterone antagonists in the rat uterus. Hum Reprod Update. 1998;4
(5):570–583.
4.
van der HorstPHWangYvan der ZeeMBurgerCWBlokLJ. Interaction between sex hormones and WNT/beta-catenin signal transduction in endometrial physiology and disease. Mol Cell Endocrinol. 2012;358
(2):176–184.
5.
GholamiKMuniandySSallehN. Progesterone downregulates oestrogen-induced expression of CFTR and SLC26A6 proteins and mRNA in rats’ uteri. J Biomed Biotechnol. 2012;2012:596084.
6.
FrolovaAIMoleyKH. Glucose transporters in the uterus: an analysis of tissue distribution and proposed physiological roles. Reproduction. 2011;142
(2):211–220.
7.
MartinLDasRMFinnCA. The inhibition by progesterone of uterine epithelial proliferation in the mouse. J Endocrinol. 1973;57
(3):549–554.
8.
MartinLFinnCATrinderG. Hypertrophy and hyperplasia in the mouse uterus after oestrogen treatment: an autoradiographic study. J Endocrinol. 1973;56
(1):133–144.
9.
MartinLFinnCATrinderG. DNA synthesis in the endometrium of progesterone-treated mice. J Endocrinol. 1973;56
(2):303–307.
10.
BartelDP. MicroRNAs: genomics, biogenesis, mechanism, and function. Cell. 2004;116
(2):281–297.
11.
VasudevanSTongYSteitzJA. Switching from repression to activation: microRNAs can up-regulate translation. Science. 2007;318
(5858):1931–1934.
12.
GonzalezGBehringerRR. Dicer is required for female reproductive tract development and fertility in the mouse. Mol Reprod Dev. 2009;76
(7):678–688.
13.
NagarajaAKAndreu-VieyraCFrancoHLDeletion of Dicer in somatic cells of the female reproductive tract causes sterility. Mol Endocrinol. 2008;22
(10):2336–2352.
14.
HongXLuenseLJMcGinnisLKNothnickWBChristensonLK. Dicer1 is essential for female fertility and normal development of the female reproductive system. Endocrinology. 2008;149
(12):6207–6212.
15.
HuSJRenGLiuJLMicroRNA expression and regulation in mouse uterus during embryo implantation. J Biol Chem. 2008;283
(34):23473–23484.
16.
LiRQiaoJWangLMicroRNA array and microarray evaluation of endometrial receptivity in patients with high serum progesterone levels on the day of hCG administration. Reprod Biol Endocrinol. 2011;9:29.
17.
KuokkanenSChenBOjalvoLBenardLSantoroNPollardJW. Genomic profiling of microRNAs and messenger RNAs reveals hormonal regulation in microRNA expression in human endometrium. Biol Reprod. 2010;82
(4):791–801.
18.
Ohlsson TeagueEMVan der HoekKHVan der HoekMBMicroRNA-regulated pathways associated with endometriosis. Mol Endocrinol. 2009;23
(2):265–275.
19.
BorenTXiongYHakamAMicroRNAs and their target messenger RNAs associated with endometrial carcinogenesis. Gynecol Oncol. 2008;110
(2):206–215.
20.
PanQLuoXToloubeydokhtiTCheginiN. The expression profile of micro-RNA in endometrium and endometriosis and the influence of ovarian steroids on their expression. Mol Hum Reprod. 2007;13
(11):797–806.
21.
BurneyROHamiltonAEAghajanovaLMicroRNA expression profiling of eutopic secretory endometrium in women with versus without endometriosis. Mol Hum Reprod. 2009;15
(10):625–631.
22.
YueLDaikokuTHouXCyclin G1 and cyclin G2 are expressed in the periimplantation mouse uterus in a cell-specific and progesterone-dependent manner: evidence for aberrant regulation with Hoxa-10 deficiency. Endocrinology. 2005;146
(5):2424–2433.
23.
KoverKLiangLAndrewsGKDeySK. Differential expression and regulation of cytokine genes in the mouse uterus. Endocrinology. 1995;136
(4):1666–1673.
24.
ChungDDasSK. Mouse primary uterine cell coculture system revisited: ovarian hormones mimic the aspects of in vivo uterine cell proliferation. Endocrinology. 2011;152
(8):3246–3258.
25.
LiRLiYKristiansenKWangJ. SOAP: short oligonucleotide alignment program. Bioinformatics. 2008;24
(5):713–714.
26.
VazCAhmadHMSharmaPAnalysis of microRNA transcriptome by deep sequencing of small RNA libraries of peripheral blood. BMC Genomics. 2010;11:288.
27.
AudicSClaverieJM. The significance of digital gene expression profiles. Genome Res. 1997;7
(10):986–995.
28.
BackesCKellerAKuentzerJGeneTrail—advanced gene set enrichment analysis. Nucleic Acids Res. 2007;35
(Web Server issue):W186–W192.
29.
WangQLuJZhangSWnt6 is essential for stromal cell proliferation during decidualization in mice. Biol Reprod. 2013;88
(1):5.
BauersachsSUlbrichSEGrossKGene expression profiling of bovine endometrium during the oestrous cycle: detection of molecular pathways involved in functional changes. J Mol Endocrinol. 2005;34
(3):889–908.
32.
KimSTMoleyKH. Regulation of facilitative glucose transporters and AKT/MAPK/PRKAA signaling via estradiol and progesterone in the mouse uterine epithelium. Biol Reprod. 2009;81
(1):188–198.
33.
WangYvan der ZeeMFoddeRBlokLJ. Wnt/Beta-catenin and sex hormone signaling in endometrial homeostasis and cancer. Oncotarget. 2010;1
(7):674–684.
34.
BehrensJJerchowBAWurteleMFunctional interaction of an axin homolog, conductin, with beta-catenin, APC, and GSK3beta. Science.
1998;280
(5363):596–599.
35.
WangYHanifi-MoghaddamPHanekampEEProgesterone inhibition of Wnt/beta-catenin signaling in normal endometrium and endometrial cancer. Clin Cancer Res. 2009;15
(18):5784–5793.
36.
SherrCJ. Cancer cell cycles. Science. 1996;274
(5293):1672–1677.
37.
ChenBPanHZhuLDengYPollardJW. Progesterone inhibits the estrogen-induced phosphoinositide 3-kinase-->AKT-->GSK-3beta-->cyclin D1-->pRB pathway to block uterine epithelial cell proliferation. Mol Endocrinol. 2005;19
(8):1978–1990.
38.
CordesKRSheehyNTWhiteMP. miR-145 and miR-143 regulate smooth muscle cell fate and plasticity. Nature. 2009;460
(7256):705–710.
39.
OstenfeldMSBramsenJBLamyP. miR-145 induces caspase-dependent and -independent cell death in urothelial cancer cell lines with targeting of an expression signature present in Ta bladder tumors. Oncogene. 2010;29
(7):1073–1084.
40.
WuYLiuSXinHUp-regulation of microRNA-145 promotes differentiation by repressing OCT4 in human endometrial adenocarcinoma cells. Cancer. 2011;117
(17):3989–3998.
41.
OzenMCreightonCJOzdemirMIttmannM. Widespread deregulation of microRNA expression in human prostate cancer. Oncogene. 2008;27
(12):1788–1793.
42.
IchimiTEnokidaHOkunoYIdentification of novel microRNA targets based on microRNA signatures in bladder cancer. Int J Cancer. 2009;125
(2):345–352.
43.
AkaoYNakagawaYNaoeT. MicroRNA-143 and -145 in colon cancer. DNA Cell Biol. 2007;26
(5):311–320.
44.
ZhuHDoughertyURobinsonVEGFR signals downregulate tumor suppressors miR-143 and miR-145 in Western diet-promoted murine colon cancer: role of G1 regulators. Mol Cancer Res. 2011;9
(7):960–975.
45.
ShaoYQuYDangSYaoBJiM. MiR-145 inhibits oral squamous cell carcinoma (OSCC) cell growth by targeting c-Myc and Cdk6. Cancer Cell Int. 2013;13
(1):51.
46.
UchimaFDEderyMIguchiTBernHA. Growth of mouse endometrial luminal epithelial cells in vitro: functional integrity of the oestrogen receptor system and failure of oestrogen to induce proliferation. J Endocrinol. 1991;128
(1):115–120.
47.
FrancoHLRubelCALargeMJEpithelial progesterone receptor exhibits pleiotropic roles in uterine development and function. FASEB J. 2012;26
(3):1218–1227.
48.
CookePSBuchananDLYoungPStromal estrogen receptors mediate mitogenic effects of estradiol on uterine epithelium. Proc Natl Acad Sci USA. 1997;94
(12):6535–6540.
49.
KuritaTYoungPBrodyJRLydonJPO’MalleyBWCunhaGR. Stromal progesterone receptors mediate the inhibitory effects of progesterone on estrogen-induced uterine epithelial cell deoxyribonucleic acid synthesis. Endocrinology. 1998;139
(11):4708–4713.
50.
LiQKannanADeMayoFJThe antiproliferative action of progesterone in uterine epithelium is mediated by Hand2. Science. 2011;331
(6019):912–916.
51.
KimJJKuritaTBulunSE. Progesterone action in endometrial cancer, endometriosis, uterine fibroids, and breast cancer. Endocr Rev. 2013;34
(1):130–162.
52.
NothnickWB. The role of micro-RNAs in the female reproductive tract. Reproduction. 2012;143
(5):559–576.
53.
CochraneDRSpoelstraNSRicherJK. The role of miRNAs in progesterone action. Mol Cell Endocrinol. 2012;357
(1-2):50–59.
54.
KlingeCM.miRNAs and estrogen action. Trends Endocrinol Metab. 2012;23
(5):223–233.
55.
ToloubeydokhtiTPanQLuoXBukulmezOCheginiN. The expression and ovarian steroid regulation of endometrial micro-RNAs. Reprod Sci. 2008;15
(10):993–1001.
BenjaminPLBurgeCBBartelPD. Conserved Seed Pairing, Often Flanked by Adenosines, Indicates that Thousands of Human Genes are MicroRNA Targets. Cell. 2005;120:15–20.
59.
XiaoweiW. miRDB: a microRNA target prediction and functional annotation database with a wiki interface. RNA. 14(6):1012–1017.
60.
BetelDWilsonMGabowA MarksDSSanderC. The microRNA.org resource: targets and expression. Nucleic Acids Res. 2008;36 (Database Issue): D149–53.
Supplementary Material
Please find the following supplemental material available below.
For Open Access articles published under a Creative Commons License, all supplemental material carries the same license as the article it is associated with.
For non-Open Access articles published, all supplemental material carries a non-exclusive license, and permission requests for re-use of supplemental material or any part of supplemental material shall be sent directly to the copyright owner as specified in the copyright notice associated with the article.
