The transcriptome of cervical ripening in human pregnancy before the onset of labor at term: Identification of novel molecular functions involved in this process

Figures & data

Table I.  Study subjects – patient demographics and clinical characteristics.

	Term no labor unripe cervix (n = 11)	Term no labor ripe cervix (n = 11)	p
Age	28 (21–38)	29 (22–37)	NS
Parity	2 (0–4)	2 (0–5)	NS
Number of prior vaginal deliveries	0 (0–0)	0 (0–4)	NS
Gestational age at delivery (weeks)	39 (38–39)	39 (37–40)	NS
Bishop score	2 (0–3)	7 (5–9)	<0.0001
Cervical dilation (cm)	0 (0–0.5)	2 (1–3.5)	<0.0001

Results expressed as median (range).NS = not significant.

Table II.  Results of Microarray Analysis.

Entrez gene	Symbol	Gene name	Fold change	FDR
6925	TCF4	Transcription factor 4	1.45	<0.00001
1462	VCAN	Versican	2.82	<0.00001
23433	RHOQ	ras homolog gene family, member Q	1.88	<0.00001
4673	NAP1L1	Nucleosome assembly protein 1-like 1	1.22	0.0001
7078	TIMP3	TIMP metallopeptidase inhibitor 3 (Sorsby fundus dystrophy, pseudoinflammatory)	2.01	0.0001
780	DDR1	dIscoidin domain receptor tyrosine kinase 1	−1.22	0.0003
54796	BNC2	Basonuclin 2	1.45	0.0003
7035	TFPI	Tissue factor pathway inhibitor (lipoprotein-associated coagulation inhibitor)	1.67	0.0005
23213	SULF1	Sulfatase 1	1.81	0.0007
800	CALD1	Caldesmon 1	1.44	0.0007
641700	ECSM2	Endothelial cell-specific molecule 2	1.85	0.0007
1289	COL5A1	Collagen, type V, alpha 1	1.69	0.0008
91624	NEXN	Nexilin (F actin binding protein)	2.34	0.0009
5069	PAPPA	Pregnancy-associated plasma protein A, pappalysin 1	1.60	0.0013
5350	PLN	Phospholamban	1.95	0.0013
9509	ADAMTS2	ADAM metallopeptidase with thrombospondin type 1 motif, 2	1.53	0.0013
389136	VGLL3	Vestigial like 3 (Drosophila)	2.32	0.0013
253827	MSRB3	Methionine sulfoxide reductase B3	1.45	0.0018
5136	PDE1A	Phosphodiesterase 1A, calmodulin-dependent	1.31	0.0019
6876	TAGLN	Transgelin	2.00	0.0022
171024	SYNPO2	Synaptopodin 2	1.51	0.0026
152137	CCDC50	Coiled-coil domain containing 50	1.44	0.0029
7168	TPM1	Tropomyosin 1 (alpha)	1.66	0.0029
2316	FLNA	Filamin A, alpha (actin binding protein 280)	1.52	0.0029
1634	DCN	Decorin	1.39	0.0039
7070	THY1	Thy-1 cell surface antigen	2.19	0.0039
274	BIN1	Bridging integrator 1	1.40	0.0044
56999	ADAMTS9	ADAM metallopeptidase with thrombospondin type 1 motif, 9	1.72	0.0044
3107	HLA-C	Major histocompatibility complex, class I, C	1.78	0.0044
2335	FN1	Fibronectin 1	1.52	0.0044
4162	MCAM	melanoma cell adhesion molecule	1.68	0.0044
10082	GPC6	glypican 6	1.50	0.0049
2099	ESR1	Estrogen receptor 1	−1.18	0.0050
1284	COL4A2	Collagen, type IV, alpha 2	1.99	0.0050
2308	FOXO1	Forkhead box O1	1.78	0.0050
633	BGN	Biglycan	1.45	0.0050
11010	GLIPR1	GLI pathogenesis-related 1 (glioma)	1.42	0.0051
348	APOE	Apolipoprotein E	1.48	0.0051
57381	RHOJ	ras homolog gene family, member J	1.54	0.0060
3535	IGL@	Immunoglobulin lambda locus	1.63	0.0061
87	ACTN1	Actinin, alpha 1	1.50	0.0066
3398	ID2	Inhibitor of DNA binding 2, dominant negative helix-loop-helix protein	1.36	0.0081
23208	SYT11	Synaptotagmin XI	1.68	0.0081
90853	SPOCD1	SPOC domain containing 1	1.92	0.0088
221981	THSD7A	Thrombospondin, type I, domain containing 7A	1.61	0.0136
30008	EFEMP2	EGF-containing fibulin-like extracellular matrix protein 2	1.68	0.0145
3992	FADS1	Fatty acid desaturase 1	1.58	0.0147
1292	COL6A2	Collagen, type VI, alpha 2	2.04	0.0147
728215	LOC728215	Similar to transmembrane protein 28	1.95	0.0202
3384	ICAM2	Intercellular adhesion molecule 2	1.84	0.0204
7837	PXDN	Peroxidasin homolog (Drosophila)	2.18	0.0205
10186	LHFP	Lipoma HMGIC fusion partner	1.45	0.0235
10979	FERMT2	Fermitin family homolog 2 (Drosophila)	1.86	0.0248
5125	PCSK5	Proprotein convertase subtilisin/kexin type 5	1.49	0.0255
3915	LAMC1	Laminin, gamma 1 (formerly LAMB2)	1.57	0.0255
1290	COL5A2	Collagen, type V, alpha 2	2.21	0.0257
93649	MYOCD	Myocardin	3.13	0.0266
55752	SEPT11	Septin 11	1.53	0.0266
1809	DPYSL3	Dihydropyrimidinase-like 3	1.78	0.0266
4499	MT1M	Metallothionein 1M	1.57	0.0266
55193	PBRM1	Polybromo 1	−1.13	0.0266
7057	THBS1	Thrombospondin 1	1.59	0.0270
79006	METRN	Meteorin, glial cell differentiation regulator	1.53	0.0282
5175	PECAM1	Platelet/endothelial cell adhesion molecule (CD31 antigen)	1.80	0.0297
116159	CYYR1	Cysteine/tyrosine-rich 1	1.51	0.0302
2534	FYN	FYN oncogene related to SRC, FGR, YES	1.47	0.0313
84937	ZNRF1	Zinc and ring finger 1	−1.25	0.0317
10579	TACC2	Transforming, acidic coiled-coil containing protein 2	−1.17	0.0320
1281	COL3A1	Collagen, type III, alpha 1 (Ehlers-Danlos syndrome type IV, autosomal dominant)	1.85	0.0322
202052	DNAJC18	DnaJ (Hsp40) homolog, subfamily C, member 18	1.25	0.0322
6614	SIGLEC1	Sialic acid binding Ig-like lectin 1, sialoadhesin	1.41	0.0322
56648	EIF5A2	Eukaryotic translation initiation factor 5A2	1.38	0.0325
23452	ANGPTL2	Angiopoietin-like 2	1.49	0.0325
3908	LAMA2	Laminin, alpha 2 (merosin, congenital muscular dystrophy)	1.73	0.0325
256435	ST6GALNAC3	ST6 (alpha-N-acetyl-neuraminyl-2,3-beta-galactosyl-1,3)-N-acetylgalactosaminide alpha-2,6-sialyltransferase 3	1.93	0.0328
83416	FCRL5	Fc receptor-like 5	2.28	0.0336
7026	NR2F2	Nuclear receptor subfamily 2, group F, member 2	1.49	0.0336
4208	MEF2C	Myocyte enhancer factor 2C	1.32	0.0336
55194	C1orf78	Chromosome 1 open reading frame 78	1.39	0.0336
8406	SRPX	Sushi-repeat-containing protein, X-linked	2.81	0.0336
649	BMP1	Bone morphogenetic protein 1	1.19	0.0343
10150	MBNL2	Muscleblind-like 2 (Drosophila)	1.29	0.0343
10205	MPZL2	Myelin protein zero-like 2	−1.60	0.0349
4256	MGP	Matrix Gla protein	1.96	0.0360
109	ADCY3	Adenylate cyclase 3	1.50	0.0363
3487	IGFBP4	Insulin-like growth factor binding protein 4	1.84	0.0371
26020	LRP10	Low density lipoprotein receptor-related protein 10	−1.23	0.0411
10439	OLFM1	Olfactomedin 1	1.59	0.0445
5793	PTPRG	Protein tyrosine phosphatase, receptor type, G	1.46	0.0453
2901	GRIK5	Glutamate receptor, ionotropic, kainate 5	−1.18	0.0453
23129	PLXND1	Plexin D1	1.49	0.0454

Genes showing differential expression between ripe versus unripe cervix in patients at term not in labor.

Genes are ranked in order of false discovery rate (FDR).

Direction of fold change denotes change in ripe cervix.

Table III.  Gene ontology analysis.

	Genes in significant list (91 genes)	Genes on array	p value*
Biological process category
Cell adhesion	12	396	<0.0001
Regulation of anatomical structure	6	71	<0.0001
Regulation of locomotion	5	67	0.005
Multicellular organismal development	25	1891	0.005
Cell motility	6	136	0.008
Phosphate transport	5	82	0.009
Localization	24	1777	0.01
Regulation of cellular component organization and biogenesis	7	207	0.01
Regulation of body fluid levels	5	110	0.02
Skin development	2	7	0.03
Blood coagulation	4	69	0.03
Molecular function category
Extracellular matrix structural constituent	10	89	2.11×10^−8
Protein binding	57	6680	0.0057
Structural molecule activity	13	648	0.0063
Integrin binding	4	43	0.0063
Glycosaminoglycan binding	5	100	0.0127
Polysaccharide binding	5	103	0.0127
Pattern binding	5	114	0.0173
Heparin binding	4	78	0.0304
Actin filament binding	3	37	0.0304
Actin binding	7	284	0.0304
Cytoskeletal protein binding	8	403	0.0495

*p values were derived using a hypergeometric distribution and were subsequently FDR corrected.

Table IV.  Results of qRT-PCR: 32 genes with differential expression between ripe versus unripe cervix in patients at term not in labor.

Symbol	Gene name	Fold change	p value
MYOCD	Myocardin	3.6	0.002*
VCAN	Versican	3.4	0.012*
THBS1	Thrombospondin 1	2.1	0.006*
FN1	Fibronectin 1	1.9	0.031*
COL4A2	Collagen, type IV, alpha 2	1.9	0.013*
COL6A2	Collagen, type VI, alpha 2	1.7	0.013*
LAMC1	Laminin, gamma 1	1.7	0.018*
LAMA2	Laminin, alpha 2	1.7	0.013*
COL5A1	Collagen, type V, alpha 1	1.6	0.037*
CTNND2	Catenin, delta 2	5.5	0.008
MMP3	Matrix metallopeptidase 3 (stromelysin 1, progelatinase)	5.3	0.042
SELE	Selectin	3.1	0.036
ADAMTS8	ADAM metallopeptidase with thrombospondin type 1 motif, 8	2.9	0.010
CTGF	Connective tissue growth factor	2.8	0.032
COL8A1	Collagen, type VIII	2.6	0.020
VCAM1	Vascular cell adhesion molecule 1	2.5	0.017
ITGB3	Integrin, beta 3 (platelet glycoprotein IIIa, antigen CD61)	2.3	0.012
SELP	Selectin P (granule membrane protein 140kDa, antigen CD62)	2.1	0.017
TIMP1	TIMP metallopeptidase inhibitor 1	2.0	0.017
VTN	Vitronectin	2.0	0.039
ITGA5	Integrin, alpha 5 (fibronectin receptor, alpha polypeptide)	2.0	0.004
COL15A1	Collagen, type XV, alpha 1	2.0	0.015
SGCE	Sarcoglycan, epsilon	1.9	0.025
COL6A1	Collagen, type VI, alpha 1	1.7	0.043
KAL1	Kallmann syndrome 1 sequence	1.7	0.012
LAMB1	Laminin, beta 1	1.6	0.012
SPARC	Secreted protein, acidic, cysteine-rich (osteonectin)	1.6	0.008
ITGB1	Integrin, beta 1 (fibronectin receptor, beta polypeptide)	1.6	0.033
ADAMTS1	ADAM metallopeptidase with thrombospondin type 1 motif, 10	1.6	0.042
CDH1	Cadherin 1, type 1, E-cadherin (epithelial)	−1.5	0.002
TGFB1	Transforming growth factor, beta 1	1.5	0.048
TIMP2	Tissue inhibitor of metalloproteinase 2	1.5	0.025

All genes in this table, except for CDH1 show higher expression levels in ripe cervix compared to unripe cervix.

Direction of fold change denotes change in ripe cervix.

*Significant by microarray and PCR analysis; fold change ≥1.5 and p < 0.05.

Figure 1.  Results of qRT-PCR assay of versican in cervical tissue in patients at term without labor: unripe cervix versus ripe cervix.

Figure 2.  Results of qRT-PCR assay of myocardin in cervical tissue in patients at term without labor: unripe cervix versus ripe cervix.