Identifying MiRNA In Pathologic Mechanisms Of Postmenopausal Osteoporosis(POMP)

BackgroundPostmenopausal osteoporosis(POMP)is one of the most common metabolic diseases affecting thousands of postmenopausal women.The clinical manifestations of POMP include back or hip pain,accompanying higher incidence of fragility fracture.Although researchers have carried out detailed research on the clinical diagnosis,treatment and pathological mechanism of POMP,especially the related cellular signaling pathways,the relevant molecular mechanisms are yet to be elucidated.Bioinformatics is an interdisciplinary tool that combines computer science and biological technology in research,analysis and interpretation of a large number of biological data.Recently,microarray and high-throughput sequencing methods have been widely used to predict therapeutic targets,including POMP.At present,the analysis of gene expression profile in peripheral blood has become one of the most effective means for the pathology and pharmacology of various diseases.Utilizing bioinformatics may be the key understand the potential pathological molecular mechanism of POMP of postmenopausal osteoporosis and multi factor related diseases in the future,which will benefit disease screening,early diagnosis and early intervention.ObejectiveTo identify and screen the key genes and their upstream targeted miRNAs that regulate the pathogenesis of POMP,and to construct the mRNA-miRNA regulatory network.MethodsWe analyzed the microarray data from GSE56116 and GSE64433 datasets to obtain the differentially expressed genes in the blood of postmenopausal women with osteoporosis and normal postmenopausal women.The functional enrichment analysis of the differentially expressed genes was carried out from three aspects of biological process,cell composition and molecular function by David database(Gene ontology,GO),and the pathway enrichment analysis was carried out by Kyoto Encyclopedia of genes and genomes(KEGG).In addition,we used TargetScan,miRWalk and miRDB databases to identify microRNAs that may target different genes,and verified these findings in induced differentiating mice Raw264.7 osteoclasts and MC3T3 osteoblasts,as well as POMP patient serum samples,through RT-PCR.ResultsThe integration analysis of the gene expression data set,a total of 847 genes were found in postmenopausal women with osteoporosis and normal postmenopausal women with significant differences in blood expression(P<0.05).Through functional enrichment analysis and literature research,the enrichment pathway of differentially expressed genes was obtained.Through the analysis of the correlation between genes and diseases,five genes VNN1,P2RX7,FASN,SLC8A1 and KL were finally screened out,which have a strong relationship with osteoporosis.Finally,the five genes and their targeted microRNAs constructed a network diagram,and the related regulatory axes were established as follows:①SLC8A1,hsa-miR-345-5p/hsa-miR-4717-3p/hsa-miR-4768-3p/hsa-miR-629-3p/hsa-miR-4793-3p② p2rx7,hsa-miR-345-5p/hsa-miR-629-3p③ KL,hsa-miR-4793-3p④VNN1,hsa-miR-629-3p⑤FASN,hsa-miR-4717-3p.The differential expression of P2RX7,SLC8A1 and miR-345-5p was found significant in the induced osteoclasts of mice and in the serum of POMP patients.ConclusionFive differential genes obtained VNN1,P2RX7,FASN,SLC8A1 and KL are all associated with postmenopausal osteoporosis(P<0.05).Among them,P2RX7,SLC8A1 genes and miRNA-345-5p are significantly correlated(P<0.05).