Screening And Clinical Significance Of Long Noncoding RNA In Adolescent Idiopathic Scoliosis

Adolescent idiopathic scoliosis(AIS)belonging to idiopathic scoliosis(IS)is the most common type,will not only affect the patient's appearance,some even affect the patient's heart and lung function.AIS pathogenesis remains unclear,with the development of gene technology,based on transcriptomics of AIS molecular typing has become a hot research how to further screening AIS incidence may be related molecule,revealing its occurrence development mechanism and search for new molecular targets for the treatment of AIS has important clinical value and prospects.Long non-coding RNA(lncRNA)is the current research focus,it can through various aspects of epigenetic,post-transcriptional level,transcriptional level of regulation of gene expression to produce.LncRNA found abnormal expression in the presence of many complex diseases such as cancer,neurodegenerative diseases,cardiovascular disease,some lncRNA has the effect of promoting or inhibiting the disease occurred.But so far,it reports on aspects of AIS related lncRNA expression and so was still very low.This research was supported through the application of RNA-Seq sequencing AIS patients and controls lncRNA expression profiling,and obtain mRNA expression profiles,and to further explore the differences LncRNA expression AIS patients,and lay the foundation for further study.The research contents are summarized as follows:(1)Expression Profiling of Peripheral Blood samples AIS patients LncRNA and mRNA.In this part of the study,we selected 10 cases of peripheral blood samples(5 normal samples,5 samples AIS)."Meaningful target gene" is defined as at least 10 peripheral blood samples have LncRNA-mRNA expression of 5 samples appears.The outcome of 10 cases in the sample,a total of 2116 LncRNA,540 Ge mRNA meet the above criteria.Evaluation: Use volcano plot and cluster analysis heat map analysis and evaluation of the data,the results are: the presence of the normal gene expression in samples and sample AIS significant difference between the expression levels of certain genes within the AIS sample and normal sample presentation significant differences.Fold difference in expression levels of more than 2 times,and after the student t test P <0.05,we assess this as a standard difference,followed by screening of the microarray data.Discovery,AIS group had 2116 LncRNAs and 540 mRNA differential expression phenomenon in which LncRNAs upregulation of 1015,down to 1101.Wherein the presence of differentially expressed LncRNA,m RNA screening,we found 64(a raised,down 63)and 26(increase 0,down 26)differentially expressed more than five times.Compared with the normal group,the sample gene AIS patients in 346 identical(?2 differential multiplier).NONHSAT137367 is the most significant down-regulated genes lncRNA(fold difference = 11.27617),and NONHSAT103134 is the most significant increase lncRNA(fold difference = 5.174644).The study found that,compared with the normal group,ARC,SET and TPM3 gene in a sample AIS patients showed significant differences.(2)peripheral blood sample at AIS and normal peripheral blood samples of 15 LncRNA which were qRT-PCR testing,validation results highly consistent with the previous results.The results show that the gene chips used in the research group of high credibility.(3)biological information prediction: the use of biological pathways KEGG methods,GO analysis and LncRNA DAVID functional annotation clustering method to the above obtained differentially expressed,mRNA functional annotation and prediction of biological information.It was found that Focal adhesion,Amoebiasis,Adherens junction,Regulation of actin cytoskeleton,Leukocyte transendothelial migration,Salivary secretion,Arginine and proline metabolism and Thyroid cancer eight specific biological pathways of potential research value.(4)we recruit four male AIS patients,four female AIS patients,four male healthy volunteers and four female healthy volunteers.After lncRNA microarray,the data is analyzed using Venn,GO,and KEGG analyses in order to select genes associated with AIS female gender preference.Results DAVID functional annotation clustering of differentially expressed mRNA analysis appears in the development of the central nervous system,skin development,transmembrane protein receptor kinase signaling pathway regulates tryptophan,lung development,metastasis and cell growth factor,indicating that AIS patients,pathological changes described above may be widespread.Useing GO analysis revealed differential expression appears LncRNA protein transcription,replication,expression of post-transcriptional gene regulation,cell differentiation and apoptosis,multiple pathologies associated with AIS evolution of metabolic protein plays an important role.Based on this,the 10 most significant differential expression of LncRNA,exactly what role to play in the onset and exacerbation AIS worth during the next step in-depth study.Venn,GO and KEGG analyses are used to discover the AIS female gender preference.A total of 134 lncRNAs are selected,which are involved with phagosome,lysosome and HTLV-1 infection,which might through negative regulation of transcription from RNA polymerase II promoter.In the present study,we first of LncRNA differentially expressed in AIS pathogenesis,and found that the associated potential virulence factors.Subsequent in-depth studies are needed to further clarify the above-mentioned downstream effector mechanisms differentially expressed.