MiR-155 affects the biological functions of trophoblastic cells through regulating cGMP-dependent kinase 1 and is involved in the mechanism of preeclampsia

doi:10.11958/20221869

Abstract

Abstract:

Objective To investigate the expression levels of miR-155 and cGMP-dependent protein kinase 1 (PKG1) in placental tissue of patients with preeclampsia, and the effect of miR-155 on the function of trophoblasts HTR-8/SVneo by inhibiting PKG1 under the mediation of nucleus factor kappa B (NF-κB). Methods Twenty placentas were collected from normal pregnant women and pre-eclampsia pregnant women who delivered by cesarean section. In vitro trophoblasts HTR-8/SVneo were cultured and divided into the NC group, the mimics group, the inhibitor group, the siRNA NC group, the PKG1 siRNA group and the siRNA+inhibitor group. Cells were treated with NF-κB inhibitor PDTC and divided into the control group, the PDTC group, the PDTC+NC group and the PDTC+mimics group. Real-time quantitative polymerase chain reaction (qPCR) was performed to detect the expression of miR-155 and PKG1 mRNA in placentas and HTR-8/SVneo cells. Western blot assay was performed to measure the level of PKG1 protein. The cell proliferation, migration and apoptosis were assessed by CCK-8 assay, Transwell assay and flow cytometry. Results The expression of miR-155 was significantly upregulated in placental tissue of the PE group, while the expression of PKG1 decreased significantly (P<0.05). The vitro experiments showed that compared with the NC group, the expression of PKG1 was significantly reduced in the miR-155 mimics group (P<0.05). The migration and proliferation ability of trophoblast was significantly weakened, the apoptotic ability was significantly enhanced (P<0.05). The expression of PKG1 was significantly increased in the miR-155 inhibitor group, the migration and proliferation ability of trophoblast was significantly enhanced, and the apoptotic ability was significantly weakened. Compared with the control group, the expression of miR-155 was significantly reduced in the PDTC group, the migration and proliferation ability of trophoblast was significantly enhanced, and the apoptotic ability was significantly weakened (P<0.05). Conclusion Results indicate that the expression of miR-155 is upregulated in preeclampsia, and can affect proliferation, migration and apoptosis of trophoblast cells by down-regulating PKG1 mediated by NF-κB.

Key words: pre-eclampsia, cell movement, cell proliferation, apoptosis, NF-kappa B, miR-155, PKG1, trophoblast

CLC Number:

R714.244

Figures/Tables 16

References 24

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基因名称	引物序列（5′→3′）	产物大小/bp
U6	上游：CTCGCTTCGGCAGCACA	96
	下游：AACGCTTCACGAATTTGCGT	96
miR-155	上游：ACACTCCAGCTGGGTTAATGCT AATCGTGATA	60
	下游：TTAATGCTAATCGTGATAGGGG	60
GAPDH	上游：TGTTCGTCATGGGTGTGAAC	154
	下游：ATGGCATGGACTGTGGTCAT	154
PKG1	上游：AACTCCACAAATGCCAGTCG	272
	下游：GCAACTGTCCTTGCCATACT	272

基因名称	引物序列（5′→3′）	产物大小/bp
U6	上游：CTCGCTTCGGCAGCACA	96
	下游：AACGCTTCACGAATTTGCGT	96
miR-155	上游：ACACTCCAGCTGGGTTAATGCT AATCGTGATA	60
	下游：TTAATGCTAATCGTGATAGGGG	60
GAPDH	上游：TGTTCGTCATGGGTGTGAAC	154
	下游：ATGGCATGGACTGTGGTCAT	154
PKG1	上游：AACTCCACAAATGCCAGTCG	272
	下游：GCAACTGTCCTTGCCATACT	272

组别	年龄/岁	BMI/（kg/m2）	SBP/mmHg
PE组	29.7±3.5	29.1±2.3	155.3±11.2
NPE组	31.0±3.1	28.3±1.9	107.3±6.2
t	1.187	1.193	16.756^＊＊
组别	DPB/mmHg	分娩孕周/周	胎儿出生体质量/kg
PE组	98.0±9.4	36.6±2.1	2.6±0.7
NPE组	67.8±6.3	37.0±1.8	3.0±0.5
t	11.957^＊＊	0.730	3.235^＊

组别	年龄/岁	BMI/（kg/m2）	SBP/mmHg
PE组	29.7±3.5	29.1±2.3	155.3±11.2
NPE组	31.0±3.1	28.3±1.9	107.3±6.2
t	1.187	1.193	16.756^＊＊
组别	DPB/mmHg	分娩孕周/周	胎儿出生体质量/kg
PE组	98.0±9.4	36.6±2.1	2.6±0.7
NPE组	67.8±6.3	37.0±1.8	3.0±0.5
t	11.957^＊＊	0.730	3.235^＊

组别	miR-155	PKG1
PE组	4.41±2.06	1.52±0.90
NPE组	2.07±0.61	4.04±2.03
t	4.782^＊＊	5.076^＊＊