The Roles And Mechanisms Of IGF2BP2 In Placental Trophoblast Invasion And Migration

BackgroundThe human placenta is an important organ for material exchange between the developing fetus and the mother.The human placenta maintains pregnancy and supports the development of the fetus by providing nutrition,gas/waste exchange,hormone regulation,and an immune barrier from the mother’s immune system,with placental trophoblast cells performing most of these functions.Eccentric invasion/migration and fusion of placenta trophoblast cells may lead to pregnancy-related diseases,such as Pre Eclampsia(PE).At present,it is generally believed that the occurrence of PE can be divided into two stages:the reduction of placental perfusion and the period of secondary maternal multiple system damage,in which the reduction of placental perfusion is considered to be one of the causes of PE.PE is a type of hypertension syndrome that occurs during pregnancy but involves multiple organs and multiple systems.It is one of the main causes of maternal and perinatal morbidity and mortality,especially Severe pre-eclampsia(sPE).PE main performance for hypertension,proteinuria and other symptoms.76,000 women and 500,000 babies worldwide die from this disease each year.Earlier prediction and diagnosis of PE and taking appropriate treatment measures to improve the survival outcomes of maternal and perinatal infants are our goals to study the pathogenesis of PE.There are several members of the IGF2BP family,among which there are Insulin like Growth Factor 2 mRNA binding protein 2(IGF2BP2).Members of this family can directly stabilize the mRNA of the target gene,thereby promoting the protein of the target gene mRNA translation.IGF2BP2 has been reported to play an important role in myogenesis,maintenance of Glioblastoma stem cells,and the onset of cancer cells Invasion and migration(mainly liver cancer,Hepatocellular carcinoma).We found in previous studies that IGF2BP2 can mediate the abnormally high expression of miR-181a-5p in plasma and placenta of sPE patients on the regulation of placental trophoblast cell invasion and migration:the abnormally highly expressed miR-181a-5p in sPE placenta may bind to the sequence at IGF2BP2 mRNA 3’-UTR 15-38 bp,results in the degradation of IGF2BP2 mRNA and protein,which reduces the invasion and migration ability of placental trophoblast.This may leads to the occurrence of sPE.These results suggest that the abnormal expression of IGF2BP2 in the placenta may be one of the causes of PE.However,the specific molecular mechanism of IGF2BP2 in placental development and trophoblast cell function remains to be studied.The purpose of this research is to clarify the role of IGF2BP2 in the invasion and migration of placental trophoblast cells and its molecular mechanism,in order to further understand the possible role of IGF2BP2 in the occurrence of PE,and to provide a theoretical basis for the diagnosis and treatment of PE.ObjectivesThrough this study,the role of IGF2BP2 in invasion and migration of placental trophoblast cells and its underlying molecular mechanism are clarified,in order to further understand the role of IGF2BP2 in the occurrence and development of PE,and provide a theoretical basis for the diagnosis and treatment of PE.Materials and Method1.Collect maternal placental tissue samples that meet the inclusion and exclusion criteria.After thorough washing,OCT embedding and liquid nitrogen cryopreservation were performed according to the needs of the experiment.2.The total protein of the sPE placenta meeting the inclusion exclusion criteria and the maternity placenta matching its gestational age were extracted,and the expression of IGF2BP2 protein was detected by Western Blot technology.3.Collect early placenta villous tissue from 8-10 weeks of gestation and use immunofluorescence technology to locate IGF2BP2 in early placenta trophoblast cell.4.HTR-8/SVneo cells were treated with IGF2BP2 siRNAs,and to investigate whether affected the invasion and migration of the HTR-8/SVneo cells by transwell invasion/migration experiments.5.1n HTR-8/SVneo cells,using specific siRNA to interfere with IGF2BP2 gene expression,the target genes that IGF2BP2 directly acts were predicted by transcriptome RNA sequencing and transcription factor prediction and literature mining.Further verification by RIP selected HMGA2 as a candidate IGF2BP2 target gene.6.HTR-8/SVneo cells were treated with HMGA2 siRNAs,and to investigate whether affected the invasion and migration of the HTR-8/SVneo cells by transwell invasion/migration experiments.Results1.IGF2BP2 expression in human sPE placentas was significantly lower than in normal placenta.2.IGF2BP2 is expressed in CT and ST cells of human first trimester placentas villous tissue.3.IGF2BP2 siRNAs suppress HTR-8/SVneo invasion and migration.4.HMGA2 is one of the direct binding substrates of IGF2BP2.5.HMGA2 siRNAs also suppress HTR-8/SVneo invasion and migration.ConclusionThis research has found that IGF2BP2 gene knockdown significantly reduced the invasion/migration ability of HTR-8/Svneo cells,which may be caused by the decrease of the stability of its downstream directly binding substrate HMGA2 mRNA.