Microarry Analysis Of Circular RNA Expression Profile And The Screening Of Potential Biomarkers For Postmenopausal Osteoporosis

BackgroundCircular RNAs?circRNAs?are a class of abundant non-coding RNA molecules.Recent studies have found that circRNAs are involved in miRNA-mediated post-transcriptional regulation and act as miRNA sponges.Previous studies have shown that many changes in miRNA expression are closely related to postmenopausal osteoporosis?PMOP?.However,the specific molecular mechanism of miRNA action has not been studied clearly,and the expression of circRNAs in PMOP is still blank.The purpose of this study is to explore the expression of circRNAs in osteoporosis and the function of human mesenchymal stem cells analysis and the potential of biomarkers.Methods1?Expression Profiles of circular RNAs in Mononuclear CellsOur circRNA microarray was used to analyze the differential expression of circRNAs in peripheral blood mononuclear cells?PBMCs?of PMOP patients and healthy controls.2?Bioinformatics analysis of differentially expressed circRNAsFive differentially expressed circRNAs were selected for RT-PCR.The circRNA-miRNA-mRNA correlation network was constructed and the GO analysis and KEGG Pathway analysis were used to carry out the preliminary study on the function of the circRNAs consistent with the chip results.3?The potential of circRNA as biomarker was assessed by the ROC curve andthe udon index.4?According to the results of the chip,five rounds of circRNAs and theirpredicted miRNAs were verified by QRT-PCR in osteogenic differentiation of human mesenchymal stem cells for 0,1,3 and 5 days.Results1?Compared with the normal control group,381 differentially expressedcircRNAs?FC?1.5,P<0.05?were found in PBMC of PMOP patients,of which 203 were up-regulated and 178 were down-regulated.2?RT-PCR results showed that the expression level of hsa_circ₀001275 wasconsistent with the results of the chip and was statistically significant.Construction of circRNA-miRNA-mRNA coexpression network,hsa_circ₀001275 target gene of miRNAs was mainly concentrated inthe biological process.KEGG Pathway analysis showed that hsa_circ₀001275 might be closely related to TNF signaling pathway.3?Hsa_circ₀001275 was associated with T-SCORE integral of bone mineraldensity,and its AUC was 0.726?95%Cl:0.585-0.868?and the Yuden index = 0.29 in the ROC curve analysis,and the diagnostic efficacy was poor.Combined ?-CROSSL diagnosis was the best:AUC= 0.786,Yuden index =0.548.4?Further verification of circRNA expression in human mesenchymal stem cells,There are four hsa_circ₀031241,hsa_circ₀000524,hsa_circ₀002082 and hsa_circ₀001275 and 8 corresponding miRNA hsa-miR-338-3p,hsa-miR-135b-5p,hsa-miR-877-3p,hsa-miR-422a,hsa-miR-422a,hsa-miR-506-3p,hsa-miR-370-3p;hsa-miR-4773 and hsa-miR-4742-3p showed consistent regular changes.hsa_circ₀000524---hsa-miR-338-3p and hsa-miR-135b-5p---targeting Runx2 and Fgfr2 pathways and hsa_circ₀001275---hsa-miR-422a---CD226 pathway may be related to the differentiation and regulation of bone marrow mesenchymal cells.ConclusionHsa_circ₀001275 is a significantly differential expression of PBMC in PMOP,which may affect the downstream gene as a miRNA sponge through the TNF signaling pathway and has certain biomarker potential.Hsa_circ₀000524---hsa-miR-338-3p and hsa-miR-135b-5p---targeting Runx2 and Fgfr2 pathways and hsa_circ₀001275--hsa-miR-422a---CD226 Pathways may play a regulatory role in human bone marrow mesenchymal cells.