The Study of MiRNA Expression Profile in Prenatal Diagnosis of Fetal Congenital Heart Disease

doi:10.12280/gjfckx.20230802

Abstract

Abstract:

Objectives: To explore the application of microRNA (miRNA) expression profile in prenatal diagnosis of fetal congenital heart disease (CHD). Methods: 30 pregnant women diagnosed with CHD by ultrasound (case group) and 10 pregnant women who required amniocentesis (control group) were collected from January 2021 to December 2022 at Tianjin Central Hospital of Gynecology Obstetrics. The amniotic fluid supernatant of the two groups of pregnant women was sequenced by Illumina sequencing platform. All miRNAs of the two groups of pregnant women were normalized and the differentially expressed miRNAs were analyzed. The miRNA with P<0.05 and |log₂ FC|>3（Fold Change, FC） were selected from the differentially expressed miRNAs for further validation by real time fluorescence quantitative polymerase chain reaction (RT-qPCR) in amniotic fluid and peripheral blood. The difference between miRNA sequencing and RT-qPCR in amniotic fluid was compared, and the miRNAs with consistent expression regulation direction in peripheral blood and amniotic fluid were selected. Results: A total of 138 differentially expressed miRNAs were found, of which 85 were up-regulated and 53 were down-regulated. Further, 15 differentially expressed miRNAs were selected, and the results of miRNA sequencing in amniotic fluid were consistent with those of RT-qPCR. There were two miRNAs with the same expression and regulation direction in peripheral blood and amniotic fluid, which were miR-222-3p and miR-189-5p, respectively. The expression of these two miRNAs in maternal blood of the case group was significantly higher than that of the control group, and the difference was statistically significant (all P<0.05). Conclusions: As a new serological marker, miRNA in maternal blood can be preliminarily applied to the screening of fetal CHD.

Key words: Heart defects, congenital, MicroRNAs, Prenatal diagnosis, Amniocentesis, Noninvasive prenatal testing, Real-time polymerase chain reaction

YANG Wei-wei, REN Chen-chun, CHANG Ying, WANG Wen-jing, JU Ming-yan, YAO Li-ying, ZHAO Xiao-min, ZHAO Dan-yang. The Study of MiRNA Expression Profile in Prenatal Diagnosis of Fetal Congenital Heart Disease[J]. Journal of International Obstetrics and Gynecology, 2024, 51(3): 342-345.

Figures/Tables 4

References 12

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miRNA	上游引物F（5′→3′）	下游引物R（5′→3′）
miR-1247-5p	GGGACCCGTCCCGTTCGTCC	CAGTGCGTGTCGTGGAGT
miR-136-3p	CTCTTAACCGCGATCCTCC	CCATCCCAAGTGACGAGCA
miR-148a-3p	GGTCAGTGCACTACAGAACTTTG	GATGATGATAAGCAAATGCTGACTGAAC
miR-154-3p	GTGGTACTTGAAGATAGGTT	TTGGTACTGAAAAATAGGTC
miR-184	AGTGCAGTGGGGTTGGTCTA	TCCAAATGGCTCATCTCCGAC
miR-19b-3p	CGTGTGCAAATCCATGCAA	CGTGTGCAAATCCATGCAA
miR-222-3p	AGGGAGGTGTCATCTCAACTGA	CTCAACTGGTGTCGTGGAGTC
miR-381-3p	TCAGACGACAACCGTCTGTG	AAAATTGAGCACCAACGGGC
miR-423-5p	CGAAGTTCCCTTTGTCATCCT	GTGCAGGGTCCGAGGTATTC
miR-4257	GAGCAGGCCAGAGGTG	ATCCAGTGCAGGGTCCGAGG
miR-432-5p	CTCGCTTCGGCAGCACA	CTTGGAGTAGGTCATTGGGT
miR-574-5p	CGCGTGAGTGTGTGTGTGTGA	AGTGCAGGGTCCGAGGTATT
miR-9-5p	AGCTTGCTGCACCTTAGTCT	TGTGTGCGGCTAGAACATCC
miR-145-3p	GGGGATTCCTGGAAATA	TGCGTGTCGTGGAGTC
miR-189-5p	CGCGGGTCGGTTAGCTC	ATCCAGTGCAGGGTCCGAGG
U6	CTCGCTTCGGCAGCACAT	AACGCTTCACGAATTTGCGT

miRNA	上游引物F（5′→3′）	下游引物R（5′→3′）
miR-1247-5p	GGGACCCGTCCCGTTCGTCC	CAGTGCGTGTCGTGGAGT
miR-136-3p	CTCTTAACCGCGATCCTCC	CCATCCCAAGTGACGAGCA
miR-148a-3p	GGTCAGTGCACTACAGAACTTTG	GATGATGATAAGCAAATGCTGACTGAAC
miR-154-3p	GTGGTACTTGAAGATAGGTT	TTGGTACTGAAAAATAGGTC
miR-184	AGTGCAGTGGGGTTGGTCTA	TCCAAATGGCTCATCTCCGAC
miR-19b-3p	CGTGTGCAAATCCATGCAA	CGTGTGCAAATCCATGCAA
miR-222-3p	AGGGAGGTGTCATCTCAACTGA	CTCAACTGGTGTCGTGGAGTC
miR-381-3p	TCAGACGACAACCGTCTGTG	AAAATTGAGCACCAACGGGC
miR-423-5p	CGAAGTTCCCTTTGTCATCCT	GTGCAGGGTCCGAGGTATTC
miR-4257	GAGCAGGCCAGAGGTG	ATCCAGTGCAGGGTCCGAGG
miR-432-5p	CTCGCTTCGGCAGCACA	CTTGGAGTAGGTCATTGGGT
miR-574-5p	CGCGTGAGTGTGTGTGTGTGA	AGTGCAGGGTCCGAGGTATT
miR-9-5p	AGCTTGCTGCACCTTAGTCT	TGTGTGCGGCTAGAACATCC
miR-145-3p	GGGGATTCCTGGAAATA	TGCGTGTCGTGGAGTC
miR-189-5p	CGCGGGTCGGTTAGCTC	ATCCAGTGCAGGGTCCGAGG
U6	CTCGCTTCGGCAGCACAT	AACGCTTCACGAATTTGCGT

miRNA	测序结果		RT-qPCR结果
miRNA	FC	P	FC	P
miR-1247-5p	6.77	<0.05	4.28	<0.05
miR-136-3p	-9.18	<0.05	-8.69	<0.05
miR-148a-3p	-7.49	<0.05	-5.63	<0.05
miR-154-3p	-9.97	<0.05	-9.56	<0.05
miR-184	-7.88	<0.05	-4.65	<0.05
miR-19b-3p	-9.42	<0.05	-6.58	<0.05
miR-222-3p	8.38	<0.05	5.15	<0.05
miR-381-3p	-9.42	<0.05	-8.73	<0.05
miR-423-5p	7.64	<0.05	6.47	<0.05
miR-4257	-9.97	<0.05	-23.43	<0.05
miR-432-5p	6.86	<0.05	7.64	<0.05
miR-574-5p	10.74	<0.05	8.33	<0.05
miR-9-5p	-5.55	<0.05	-9.34	<0.05
miR-145-3p	7.20	<0.05	6.43	<0.05
miR-189-5p	7.07	<0.05	6.49	<0.05

miRNA	测序结果		RT-qPCR结果
miRNA	FC	P	FC	P
miR-1247-5p	6.77	<0.05	4.28	<0.05
miR-136-3p	-9.18	<0.05	-8.69	<0.05
miR-148a-3p	-7.49	<0.05	-5.63	<0.05
miR-154-3p	-9.97	<0.05	-9.56	<0.05
miR-184	-7.88	<0.05	-4.65	<0.05
miR-19b-3p	-9.42	<0.05	-6.58	<0.05
miR-222-3p	8.38	<0.05	5.15	<0.05
miR-381-3p	-9.42	<0.05	-8.73	<0.05
miR-423-5p	7.64	<0.05	6.47	<0.05
miR-4257	-9.97	<0.05	-23.43	<0.05
miR-432-5p	6.86	<0.05	7.64	<0.05
miR-574-5p	10.74	<0.05	8.33	<0.05
miR-9-5p	-5.55	<0.05	-9.34	<0.05
miR-145-3p	7.20	<0.05	6.43	<0.05
miR-189-5p	7.07	<0.05	6.49	<0.05

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