Differential Expression Analysis Of Placenta-related Long Noncoding RNAs And The Biological Function Of MIR210HG In Preeclampsia

Objectives: In order to detect the changes of RNA in placental tissues of preeclampsia(PE),we investigated the differential expression of long noncoding RNAs(lncRNAs).In this study,the mechanism of lncRNA MIR210 HG,highly expressed in the placenta of preeclampsia,was explored in order to provide a new perspective for the occurrence and development of PE.Methods:(1)The differential expression of lncRNAs in PE and normal pregnancy placental tissues were analyzed by RNA-seq technique.(2)The expression levels of MIR210 HG in placental tissues were further verified by Real-Time PCR,and the correlation between the expression level of MIR210 HG and the clinical indicators of the subjects in the two groups was analyzed.(3)Through cell proliferation and migration experiments in vitro,the effect of MIR210 HG on HTR8/Svneo cell line and Eahy cell line was preliminarily investigated.(4)The possible pathways of MIR210 HG were predicted by bioinformatics analysis.(5)Down-regulating MIR210 HG by small interfering RNA(si RNA),the changes of key genes of Renin-Angiotensin System in HTR8/ SVneo cell line were detected by RT-PCR and Western blot.Results:(1)A total of 26 significantly differentially expressed lncRNAs were found in the RNA-sequencing analysis,among which 21 were up-regulated and 5 were down-regulated.(2)RT-PCR results showed that the expression level of MIR210 HG in PE placental tissues was up-regulated.Systolic and diastolic blood pressure had positive linear correlation with MIR210 HG expression levels,while neonatal weight had a negative linear correlation with MIR210 HG expression levels(P<0.05).However,there was no significant correlation between the expression level of MIR210 HG and BMI(P=0.0582),serum creatinine level(P= 0.6522),albumin level(P= 0.6522)and platelet level(P= 0.2000)before delivery.(3)In vitro experiments,the proliferation and migration of HTR8/SVneo and Eahy were both enhanced after MIR210 HG knockdown by si RNA(P<0.05).(4)The MIR210 HG pathway enrichment analysis showed that Gene Ontology(GO)analysis was enriched in 27 pathways,Kyoto Encyclopedia of Gene and Genomes(KEGG)in 8 pathways,and Bio Carta in 8 pathways.The prediction results of RNA binding with MIR210 HG showed that 38 RNAs interacted with MIR210 HG,including 24 m RNAs and 3 r RNAs.(5)After MIR210 HG knockdown,the expression of Vascular endothelial growth factor(VEGF)was increased at both m RNA and protein levels(P<0.05),while the expressions of Angiotensin I converting enzyme 1(ACE1),Renin(REN),ATPase H+ transporting accessory protein 2(ATP6AP2),Angiotensin II receptor type 1(AGTR1)and Angiotensinogen(AGT)genes in HTR8/SVneo cells were not significantly changed.Conclusions:(1)lncRNA MIR210 HG was found to be up-regulated in PE placental tissues.(2)The expression level of MIR210 HG might be correlated with the severity of PE.(3)As a negative regulator,MIR210 HG played a role in proliferation and migration of trophoblast cell and endothelial cell.(4)MIR210HG might be involved in the biological behavior of trophoblast cell by affecting the expression level of VEGF in RAS key gene.