Background
Methods
Oocyte donors and ovarian stimulation
Luteal-phase support and tissue collection
RNA preparation and miRNA analysis
Statistical analysis
Results
Demographic characteristics
Characteristics | Group IIa (no support) | Group IIb (P support) | Group IIc (P + E support) | P |
---|---|---|---|---|
N
| 3 | 3 | 3 | |
Age (yr)
| 25.7 ± 3.2 | 23.6 ± 0.8 | 22.8 ± 1.2 | 0.494 |
BMI (kg/m
2
)
| 23.3 ± 1.4 | 21.6 ± 1.8 | 20.2 ± 1.2 | 0.096 |
Day 2 FSH (mIU/ml)
| 4.5 ± 0.9 | 5.6 ± 1.1 | 4.0 ± 0.3 | 0.178 |
Day 2 LH (mIU/ml)
| 2.4 ± 0.8 | 4.0 ± 1.3 | 5.2 ± 0.2 | 0.507 |
Day 2 E2 (pg/ml)
| 36.7 ± 11.6 | 34.5 ± 12.5 | 20.5 ± 3.5 | 0.646 |
Gonadotropins used (IU)
| 2850 ± 525 | 2400 ± 645 | 2625 ± 675 | 0.387 |
Peak E2 level (pg/ml)
| 1928 ± 100.0 | 2514 ± 400 | 2625 ± 480 | 0.563 |
Days of stimulation
| 10.3 ± 1.1 | 9.3 ± 1.2 | 10.1 ± 0.7 | 0.588 |
No. of oocytes
| 14.5 ± 5 | 18.4 ± 3 | 16.0 ± 4 | 0.398 |
MiRNA profiles and comparisons between groups
Venn diagram analysis of differentially expressed miRNA genes
Validation analysis
MiRNA and target genes
miRNA | P vs. no | P + E vs. no | P + E vs. P | Predicted target genes |
---|---|---|---|---|
hsa-miR-335
|
↑↑
| IMP2,CD79B,WWP1,AP3S1,HOXD8,MAX,SP1,MAP2,MAK3,STARD7,CAP350,PANK2,SRPR,PPP6C,LASS5,ATP1B1 | ||
hsa-miR-346
|
↑
|
↑
| IMP1,EIF3S1,BCL6,ABCC12,LIF,FSTL4,KGFLP2,KRAS,RAF7,FGF7,TMEM28,IGSF4B,PPP1R9B,COL2A1,HCG3, CALN1,HBP1,SF1 | |
hsa-miR-448
|
↑
| DOCK9,PPM2C,NTF3,CAP1,MAP2K6,ITM1,PRKAR2B,PAPPA,CDC2L6,CNTN4,IGF1R,SOCS5,CLK1,HOXA11,WWP1,FOXJ3,WDR22,MPPED2,ADD1,PRKA2 | ||
hsa-miR-504
|
↑
| DCX,ATP1B4,IL1RAPL1,MNT,KLF13,PRKAR2A,IL16,LIF,FXR2,NRF1,CAMK2G,MMD,LOC284296,DND1,CNTFR, SORT1,NFIX | ||
hsa-miR-512-5p
|
↑↑
|
↓
| PIK3R1,CTNNB1,EMX2,SOX21,RIPK5,MBD6,SRPK, VNN3,ERP29,PHF15,FBXW11,LOC285074,MAP1A,CHD9 | |
hsa-miR-520 g
|
↑↑
| ETF1,CAMK2N1,NLK,TNFSF11,CNR1,EFTUD1,HMGB3,FBN2,ENC1,MARK4,TFEB,TNFSF12,PRKAR2A,TNKS1BP1,EIF4E,PPP3CA,IMP1,MAP3K14,TMTC2,TTN,GTF2IRD2,PTK2B,DNAJB5,TNRC6A,VEGF,EIF4G2,FOXO1A,MAP3K9 | ||
hsa-miR-204
|
↑
| RUNX2,SOX4,NRBF2,MAP1LC3B,CDC42,ATP2B1,AKAP1,MAP3K3,CENTD1,IGF2R,NTRK2,TGFBR2,AP3M1, | ||
NEUROG1,P53CSV,TCF7L1,CDH2, CDC25B,TCF12,ELF2 | ||||
hsa-miR-369-3p
|
↓
| TCF8,PKIA,TLN1,CHD7,NCK2,CD2AP,CDC2L6,ELMOD1,CCNE2,FOXG1B,HOXB3,ADAMTS19,GIT2,ADAMTS3,TCF12, SRPK2,ADAMTS6,MAP2,ADAM10,FOXO1A,VEGFC | ||
hsa-miR-328
|
↑
| AK6,ITGA5,PRX,IGSF4C,MAX,SOX11,PTPN9,DPH2, HIST1H4D,USP37,VSIG4,RPP14,SF4,ULK2,FGD1,PLAG1 | ||
hsa-miR-186
|
↑
| APLP2,ITGA6,RPS6KB1,CDC42,PRDM10,IGSF11,EFCBP1,TCF20,CAST,LMBR1,TMED2,TGFBR2,ICMT,IL2,CCNT2,HOXB8,PAK7,FOXD1,PTGES3,MAP3K2,VEGF,COL3A1, SRPK2,MAPKAP1,C16orf52,MAP2 | ||
hsa-miR-517a
|
↑
| AMMECR1,ACACA,NPAS4,BSN,HNRPU,PTK2B,CDKN2A,CBLN2,RAPGEFL1,LOC201895,FOXJ3,PHF13,TMCC1 | ||
hsa-miR-365
|
↓↓
| EIF4E3,MAP2K7,LAMP2,ENTPD7,PCNP,ADAMTS6, COL7A1,PPP5C,REV3L,PTGDR,KCNH2,RBM12,PKD2L2 | ||
hsa-miR-221
|
↓↓
| CDC2L6,TIMP3,EIF4E3,NTF3,IMP2,HTLF,CDV3,NL,EIF5A2,NRK,PAK1,CDKN1C,FAT2,LIFR,TMCC1,MAP3K10,VGLL4,FAM13A1,TCF12,HOXC10,MAPK10,HMGCLL1,ADAM11,CD4,CTCF | ||
hsa-miR-495
|
↓↓
| MAPK6,CDK6,EML4,ILF3,PTK9,PRR7,HBEGF,HOMER1,MARK3,SP4,TGFB2,LHX2,HOXC6,PRKX,AP3M1,SOX9,GMFB,HMGCLL1,FOXO3A,EDG3,NKRF,HOXB9,TIMP2, IGSF4,CD164,TNFRSF21 | ||
hsa-miR-146a
|
↓
|
↑
| FBXL10,IRAK1,TRAF6,CD79B,SP8,FLJ33814,SFRS6,NPAS4,CXorf22,EIF4G2,MMP16,USP3,KCTD15,SMAD4,LOC440944,SEC23IP,BCORL1,TM6SF2,DLGA1 | |
hsa-miR-99a
|
↓
|
↓
| EPDR1,FZD8,HS3ST2,EIF2C2,HS3ST3B1,FGFR3,BAZ2A,MBNL1,CYP26B1,KBTBD8,SMARCA5,FRAP1,ZZEF1,ICMT,C4orf16,ADCY1,MTMR3,CTDSPL,HOXA,RAVER2, INSM1,TRIB2,SLC44A1 | |
hsa-miR-181c,d
|
↑↑
| ETF1,COL16A1,NLK,TNFSF11,MAP3K3,MAP2K1,ITGA3,TCERG1,MAPT,MAPK1,MAP1B,CDH13,ITGB8,PCGF2,ADAMTS18,LMBRD2,MMP14,CD163,LIF,ADAMTS6, TNFRSF11B,CDC42BPA | ||
hsa-miR-200b
|
↓
| TCF8,NTF3,CYLN2,HMGB3,PRKAR2B,MPP5,GIT2,MAP4K3,FLJ21103,E2F3,CSNK1G3,MMD2,ZNF53, EIF5B,ERRFI1 | ||
hsa-miR-196b
|
↓
| IMP1,CDYL,COL14A1,SSR1,IMP3,CDV3,CALM3,COL24A1,CDKN1B,ELF4,HOXC8,HMGA2,HOXA5,MAP4K3,PARP6,COL3A1,HOXA1,TNFSF12,COL1A2, HOXA7,HOXB6 |
Symbol | GrpsIIa-I [ILL-FC] | GrpsIIc-IIa [ILL-FC] | Source | Symbol | Entrez gene name | Pathway (enrichment) | P Value | FDR |
---|---|---|---|---|---|---|---|---|
miR-223-3p (GUCAGUU)
| 1.389 | 1.111 | 1 | VIM | vimentin | |||
miR-223-3p (GUCAGUU)
| 1.389 | 1.111 | 1,2 | RHOB | ras homolog family member B | |||
miR-223-3p (GUCAGUU)
| 1.389 | 1.111 | 1 | IRS1 | insulin receptor substrate 1 | |||
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 2,3 | TUBB2A | tubulin, beta 2A class IIa | |||
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 1,2,3,4 | SPARC | secreted protein, acidic, cysteine-rich (osteonectin) | extracellular matrix | 1.29E-07 | 4.44E-06 |
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 1,2,3 | PIK3R1 | phosphoinositide-3-kinase, regulatory subunit 1 (alpha) | |||
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 2,3 | MYBL2 | v-myb myeloblastosis viral oncogene homolog (avian)-like 2 | |||
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 1,2 | COL5A3 | collagen, type V, alpha 3 | extracellular matrix | 1.29E-07 | 4.44E-06 |
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 32,3 | COL5A2 | collagen, type V, alpha 2 | extracellular matrix | 1.29E-07 | 4.44E-06 |
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 1,2,4 | COL4A1 | collagen, type IV, alpha 1 | extracellular matrix | 1.29E-07 | 4.44E-06 |
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 1,2,3,4 | COL1A2 | collagen, type I, alpha 2 | extracellular matrix | 1.29E-07 | 4.44E-06 |
miR-29b-3p (AGCACCA)
| 1.375 | 1.068 | 1,2,4 | COL15A1 | collagen, type XV, alpha 1 | extracellular matrix | 1.29E-07 | 4.44E-06 |
miR-9-5p (CUUUGGU)
| 2.104 | −1.802 | 1 | NFKB1 | nuclear factor of kappa light polypeptide gene enhancer in B-cells 1 | |||
miR-9-5p (CUUUGGU)
| 2.104 | −1.802 | 1,2 | FOXO1 | forkhead box O1 | |||
miR-9-5p (CUUUGGU)
| 2.104 | −1.802 | 1,2 | FOXG1 | forkhead box G1 | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-9-5p (CUUUGGU)
| 2.104 | −1.802 | 1,2,3 | CDH1 | cadherin 1, type 1, E-cadherin (epithelial) | |||
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,4 | TRA@ | T cell receptor alpha locus | |||
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2 | TIMP3 | TIMP metallopeptidase inhibitor 3 | |||
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2 | NOTCH4 | notch 4 | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2 |
KRAS
| v-Ki-ras2 Kirsten rat sarcoma viral oncogene homolog | response to steroid hormone stimulus | 6.24E-07 | 4.65E-05 |
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2,4 | HOXA11 | homeobox A11 | |||
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2 | GATA6 | GATA binding protein 6 | |||
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2 | ESR1 | estrogen receptor 1 | response to steroid hormone stimulus | 6.24E-07 | 4.65E-05 |
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2 | CDKN1B | cyclin-dependent kinase inhibitor 1B (p27, Kip1) | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-181a-5p (ACAUUCA)
| 1.376 | −1.24 | 1,2,3,4 |
BCL2
| B-cell CLL/lymphoma 2 | response to steroid hormone stimulus | 6.24E-07 | 4.65E-05 |
miR-196a-5p (AGGUAGU)
| 1.092 | −1.342 | 1 | IKBKB | inhibitor of kappa light polypeptide gene enhancer in B-cells, kinase beta | |||
miR-196a-5p (AGGUAGU)
| 1.092 | −1.342 | 1,2,4 | HOXC8 | homeobox C8 | |||
miR-196a-5p (AGGUAGU)
| 1.092 | −1.342 | 1,3 | ANXA1 | annexin A1 | |||
miR-99a-5p (ACCCGUA)
| 1.427 | −1.48 | 1,2,3 | MTOR | mechanistic target of rapamycin (serine/threonine kinase) | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-99a-5p (ACCCGUA)
| 1.427 | −1.48 | 1,2,3 | IGF1R | insulin-like growth factor 1 receptor | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-99a-5p (ACCCGUA)
| 1.427 | −1.48 | 1,2,3 | FGFR3 | fibroblast growth factor receptor 3 | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-128 (CACAGUG)
| 1.184 | −1.345 | 1,2 | TXNIP | thioredoxin interacting protein | response to steroid hormone stimulus | 6.24E-07 | 4.65E-05 |
miR-128 (CACAGUG)
| 1.184 | −1.345 | 2,3 |
TGFBR1
| transforming growth factor, beta receptor 1 | response to steroid hormone stimulus | 6.24E-07 | 4.65E-05 |
miR-128 (CACAGUG)
| 1.184 | −1.345 | 1,2 | LDLR | low density lipoprotein receptor | response to steroid hormone stimulus | 6.24E-07 | 4.65E-05 |
miR-128 (CACAGUG)
| 1.184 | −1.345 | 1,2 | E2F3 | E2F transcription factor 3 | positive regulation of cell proliferation | 1.21E-08 | 1.67E-06 |
miR-128 (CACAGUG)
| 1.184 | −1.345 | 1,2 | ADORA2B | adenosine A2b receptor |