LINC01291 Regulates Activity Of Trophoblast Cell And Affects Recurrent Spontaneous Abortion

Recurrent spontaneous abortion(RSA)refers to two or more spontaneous abortions with multiple and complex causes,half of which are unclear,and is called unexplained recurrent spontaneous abortion(URSA).URSA is a common and difficult clinical disease,and its pathogenesis covers multiple hypotheses such as anatomical abnormalities and immune system disorders.The specific pathogenesis remains to be further explored.Long non coding RNA(lncRNA)refers to RNA fragments that are longer than 200 bp and do not have the ability to encode.Originally considered as "noise" of transcription,it has no biological function.With further research,it has gradually been found that it can be divided into antisense long chain non coding RNA(Anti sense lncRNA),intron non coding RNA(Intronic Non coding RNA),and large inter genic non coding RNA(LincRNA).Through follow-up research,it has gradually been found that it has important biological functions.LncRNA can play a role through transcriptional interference,chromatin remodeling,regulating variable splicing patterns,producing endogenous siRNA,acting as a precursor of miRNA,affecting mRNA stability,protein activity,protein transport,and protein degradation,and acting as endogenous competitive RNA(ceRNA).MicroRNA(miRNA)refers to a nucleotide sequence with a length of about 20 bp,which can undergo complete and/or incomplete complementary pairing with other nucleotide sequences such as lncRNA and mRNA.When miRNA binds to mRNA,the translation ability of downstream mRNA is blocked and subsequently degraded,thereby regulating protein expression.The ceRNA mechanism is one of the classic modes of action of lncRNA,which can act as a molecular sponge to adsorb miRNA and reduce the binding of miRNA to downstream mRNA.In previous studies,we found that compared with normal abortion patients,the expression of human chromosome segregation like gene-1(CSE1L)in villous of patients with recurrent abortion decreased.By changing the expression of CSE1L in trophoblasts,we found that it had a significant impact on the proliferation,invasion,and migration ability of cells.In order to explore its mechanism of action,the research team conducted high-throughput sequencing of three groups of HTR-8/SVneo after knocking down CSE1L and normal HTR-8/SVneo.The sequencing results showed that there was the most significant difference in the overexpression of LINC01291 in HTR-8/SVneo after knocking down CSE1L compared to the control group.After consulting the database,we learned that LINC01291 is located on human chromosome 2,with a length of 494 bp,and its subcellular location is cytoplasm.Subsequently,we established a knockdown stable transfection strain of LINC01291 in HTR-8/SVeno cells,and verified through CCK-8,cell scratch and transwell tests that compared with normal HTR-8/SVeno cells,LINC01291 knockdown reduced their proliferation,migration,and invasion abilities.Subsequently,we collected clinical samples from RSA patients and normal abortion patients for testing,and found that the expression of LINC01291 in chorionic villi of RSA patients was significantly higher than that of the normal group(P<0.001);Although the decidual tissue species LINC01291 showed an upward trend in RSA patients,there was no significant statistical difference(P>0.05).We have found a total of 30 miRNAs that may bind to LIINC01291,and 24120 target proteins of miRNAs through the method of biogenic analysis.Through GO analysis,we found that the functions of these proteins were mainly enriched in cell biological processes,cell structural composition,and acting as binding proteins;KEGG suggests that the target protein is mainly enriched in cell signal transduction and cell metabolism pathways,with the most significant enrichment in phosphatidylinositol metabolism and phosphatidylinositol signal transduction pathways.In order to further explore the interaction between their target proteins,we analyzed the top 5 miRNAs with the strongest binding ability of LINC01291 and the 10 proteins with the strongest binding ability of these 5 miRNAs,and obtained their protein interaction patterns.This topic screened and verified the high expression of LINC01291 in RSA and its regulation of trophoblast cell function through methods such as high-throughput sequencing,basic research,clinical sample analysis,and biographical analysis,and analyzed its possible mechanism of action,enriching the pathogenesis of RSA and providing new ideas for clinical treatment.