LncRNA AK002210 Participates In The Pathogenesis Of Preeclampsia By Regulating Trophoblast Invasion

Objective:preeclampsia(PE)is a common idiopathic disease during pregnancy,which mainly occurs 20 weeks after pregnancy.Its clinical manifestations are mainly hypertension,proteinuria,edema and so on.Preeclampsia is the main inducing factor of fetal and neonatal death.It is very important to understand the pathogenesis of preeclampsia for its prevention and treatment.The pathogenesis of early-onset preeclampsia is very complex,involving a variety of physiological processes,but the exact cause and pathogenesis are still uncertain.Nevertheless,it is certain that trophoblasts play an important role in the development of preeclampsia.LncRNA is related to the occurrence and development of many diseases.Studies have shown that lncRNA is involved in many cell biological processes,including cell proliferation,cell migration,cell apoptosis,X-chromosome inactivation,gene imprinting and stem cell transformation.More and more studies show that lncRNA also plays an important role in preeclampsia.MicroRNA is a kind of endogenous single stranded noncoding RNA with a length of 22-25 NT,which regulates gene expression by binding to the untranslated region(3’UTR)of the target gene.Post transcriptional regulation of gene expression,such as translation inhibition or mRNA cleavage,can silence target gene expression.The hypothesis of competing endogenous RNA(ceRNA)reveals a new mechanism of RNA interaction.CeRNA regulates the expression of multiple transcripts by binding different transcriptional response elements(MRE).Because ceRNA can compete for miRNA binding,which affects gene silencing caused by miRNA binding to mRNA,it reveals the existence of an lncRNA miRNA regulatory pathway.The purpose of this study is to elucidate the molecular mechanism of lncRNA AK002210 regulating the proliferation,invasion and apoptosis of trophoblasts through the mechanism of ceRNA.Methods:We collected serum samples from patients with early-onset preeclampsia(EOPE)and gestational age matched healthy pregnant women,as well as placenta tissue samples after delivery.According to the references,we selected the differential expression of lncRNA AK002210,and qPCR confirmed that there was differential expression of lncRNA in EOPE group and normal group.We cultured the immortalized trophoblastic cell line HTR8/svneo and the overexpression or interference of lentivirus against AK002210.Apoptosis of trophoblast was detected by annexin V/PI,proliferation of trophoblast was detected by MTT and EDU staining,migration and invasion of trophoblast were detected by Transwell experiment,and changes of AK002210 expression on trophoblast function were studied by flow cytometry.The potential binding MircroRNA of lncRNA AK002210 was predicted by bio informatics analysis.Luciferase experiment and RNA pull down experiment were used to prove whether AK002210 can directly bind miR-590-3p.The rescue experiment further proved the interaction between AK002210 and miR-590-3p.The target gene of miR-590-3p was also predicted and confirmed by bioinformatics analysis,luciferase experiment and RNA pull down experiment.Pearson analyzed the correlation between AK002210 and NAIP,AK002210 and miR-590-3p.Immunofluorescence and Western blot were used to detect protein expression.Results:AK002210 overexpression can significantly enhance the proliferation,migration,invasion and inhibition of apoptosis of trophoblasts and inhibit oxidative stress in cells.AK002210 interference can inhibit the proliferation,migration,invasion and apoptosis of trophoblasts,and enhance the oxidative stress in cells.Bioinformatics analysis predicted that miR-590-3p might be the binding gene of AK002210.Luciferase experiment and RNA pull down experiment showed that AK002210 could directly bind miR-590-3p.Rescue experiment confirmed that miR590-3p could reverse the effect of AK002210.FISH experiment confirmed the colocation of AK002210 and miR-590-3p.Similarly,bioinformatics analysis,luciferase experiment and RNA pull down experiment confirmed that miR-590-3p could target NAIP.Pearson analysis indicated that AK002210 was negatively correlated with miR-590 expression,which belonged to ceRNA relationship.AK002210 interference can inhibit the phosphorylation of ERK1/2 and the expression of MMP-2 through miR-590-3p,promote the expression of Bax and Capsase3,and inhibit the expression of Bcl-2.Conclusion:AK002210 can enhance the expression of NAIP and regulate the downstream apoptotic signaling pathway,and participate in the regulation of ERK1/2/MMP-2 signaling pathway,playing an important role in the regulation of trophoblast function.AK002210 is expected to be a potential and valuable diagnostic molecule for PE.