The Role And Mechanism Of Placental Mir-141-3p In Non-diabetic Macrosomia

ObjectiveAs one of the most common perinatal complications of pregnancy,non-diabetic macrosomia(NDFMS)has a serious impact on the long-term health of the baby.Many studies showed microRNAs(miRNAs)could regulate the placental development,yet the role and mechanism in NDFMS remains unclear.Our aim was to identity the key aberrantly expressed miRNAin the placental tissue of NDFMS,and to study the role and mechanism of the miRN A in the development of NDFMS.Methods1.We screened out the key miRNAs that abnormal expressed in NDFMS placenta through miRN A microarray using case-control study.We performed quantitative RT-PCR(qRT-PCR)analysis to validate the key miRN A and done the association analys is between the key miRN A and birthweight of fetus.2.Using HTR-8/SVneo cell as a model,westudied the effect of key miRNA on cells to explore the molecular mechanism of its involvement in NDFMS.We constructed the in vitro cell model of enhanced or knocked down expression of key miRNAby transient transfection with mimics or inhibitors.CCK8,Transwell test and flow cytometry assays were used to evaluate the cellular function.The bioinformatics software such as TargetScan,miRDB,RNA22,PicTar,PITA and others were used to predict the potential target genes of the key miRNA.Then we verified their expression,and studied the function by trans fection of the overexpress ion plasmid.3.MiR-141-3p agomir was injected into pregnant mice(early pregnancy or late pregnancy)via tail vein to construct miR-141-3p overexpression model in placental tissues.The birth weight of offspring was observed in different groups.Results1.We identified 12 differentially expressed miRNAs,including 10 up-regulated(miR-331-3p,miR-204-5p,miR-106b-5p,miR-370-3p,miR-205-5p,miR-141-3p,miR-126-3p,miR-424-5p,miR-194-5p,miR-411-5p)and 2 down-regulated(miR-1291?miR-1283).MiR-331-3p,miR-370-3p,miR-141-3p,miR-126-3p,miR-424-5p,miR-411-5p expression levels were confirmed differently between NDFMS and controls by qRT-PCR,and P values were 0.047,0.034,<0.001,0.023,0.031,0.040.Considering P value and the fold change of expression,miR-141-3p was determined as the key miRNA with the most significant difference.The correlation analysis showed fetal birth weight increased along with the increase of miR-141-3p expression(P=0.019,r2=0.101).2.MiR-141-3p could promote the proliferation,migration and invasion ability of HTR-8/SVneo cells,and changed the expression level of potential target gene PLAG1(significantly down-regulted compared with control).Overexpression of PLAG1 could reverse the effect of cell proliferation and invasion ability caused by miR-141-3p overexpression.3.No significantly difference of fetal birth weights was observed between control group and treated group with miR-141-3p agomir in early pregnancy.Whereas the fetal birth weights of treated group with miR-141-3p agomir in late pregnancy were significantly higher than control group.ConclusionThis study revealed miR-141-3p could increase the proliferation of placenta to participate in the occurrence and development of NDFMS through regulating PLAG1 expression.