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Wang et al. J Transl Genet Genom 2023;7:126-40 https://dx.doi.org/10.20517/jtgg.2023.07 Page 130

Table 1. Primer sequences used for qRT-PCR
Genes Forward primers (5′-3′) Reverse sequence (5′-3′)
lncRNA LncADAM9 TACATACAATGGTCAGATGGCAAA GCCTTGATGGGAACTGCTGA
LncPCDH10 TGAGGGAACAGGTTCTCTATGGT TGTGTGGTTCTGATGTGTCTCCTAC
lncUSP46 CGGGGACGATCAGTCACATAAG CGTGCTGGGATTGGTCATAGTT
mRNA ADAM9 GCCACTGGGAATGCTTTGT GCAGTATTCATTTCATTGTATGTAGGT
PCDH10 TGTCCAACGAGACTAAACACCAG CTATCTCCCTGTTCACTGTCTCCAT
USP46 AGAAGCCCAGAAAAGGATGAGG GAACGACCACCGCAACCAA
CADM1 CCACAGGAAAGTTACACCACCAT CACCTCCGATTTGCCTTTTAG
CD274 CAATGTGACCAGCACACTGAGA CAGTGCTACACCAAGGCATAATAAG
CLDN7 AGGGGCTGTGGATGGACTG GCCTTCTTCACTTTGTCGTCTC
CNTN1 ATAGAAGTCCCAATCCCCAGAGA GGGTGCACCTGAAATTTTGACT
HLA-DPB1 CGGAGTAAGACATTGACGGG CTCGTTGAACTTTCTTGCTCCT
ICAM2 TCCAGGATCGGATGAGAAGGTA GGAGATGTTTGAGACCAAGTAATG
ITGAL AGCAGCAAGCATTTCCACCT GGATTCCATTCGGGACACCT
NRXN2 AGAACCAGGCATCCCTCCCT AGATGCCGGCTTCCCTCAG
SELL ACTGGGATTGAAGAAACCACCTG CCAGGGGATGGCTACAGTTCA
SIGLEC1 CTCTGTCACCTTCAACAGCCAGA TGGGGGCATACTTCACTTGGA
CADM3 ACCCTCAATGTTAATGACCCCA AGGAAGACAATGAAAGCCACG
MPZL1 GCTTGGGCTCTTGACAGCT GTCAACCCGCCAGTCGTA
SPN CCTTGGGGTGCTGGTGGTA GAGGTAGGGCTGAGGTCTGGTC
CLDN10 TCATACTGTCAGGGCTGTGCTC CTCCTGCCCATCCAATAAACA
ICAM3 CGAGTCCTGTATGGTCCCAAAAT CGTGTCTCGTTTTATCTTTCCATT
ITGB7 CTCTGCGGAGGCTTTGGT ACCATAGTAGCCGTCCAAGCA
GAPDH TTGGCTACAGCAACAGGGTG GGGGAGATTCAGTGTGGTGG

antibodies (Abcam, Cambridge, MA), which are against eight proteins that are involved in the ECM-A
pathway and are differentially expressed in sPTB, were diluted to 1:1,000, and the peroxidase-conjugated
secondary antibody (Abcom), to 1:5,000. The bands were detected by chemiluminescence signals by using
ECL Western Blotting Substrate (Thermo Fisher) and scanned to produce digital images. GAPDH served as
an internal control.

Statistical analysis
For quantification of RNAs, ∆Ct = Cttarget - Ctreference was computed for each sample. ∆Ct values,
surrogates for RNA abundance in each sample, were compared between groups using an unpaired, two-
tailed t-test. Differences in DEP were considered significant at P < 0.05. The final result was finally reported
as relative expression by setting the expression value in the control group as “1”. The expression value in the
case group was calculated in relation to the control group. All data were given in terms of relative
expression of mean ± SD. The data were subjected to one-way ANOVA, followed by an unpaired, two-tailed
t-test.

RESULTS
Human placental incRNAs associated with ECM-A pathway
The original transcriptomic profile with identification of differentially expressed lncRNAs was deposited in
Gene Expression Omnibus with an accession number GSE 50879 (Available from: http://
www.ncbi.nlm.nih.gov/geo/info/linking.html). Transcriptomic analysis of chorionic villi from 40 human
placenta samples (10 from each subgroup) showed that significant up-regulation of lncADAM9 in pPROM
and sPTL as compared to samples from either FTB or PROM [Figure 1].