The Differently Expressed MicroRNAs In Serum Between Osteoporosis Patients And Normal People

Objective:miRNA (microRNA), endogenous short length of non-coding RNA, is a class of post-transcriptional gene expression regulators to influence cell differentiation, proliferation, and apoptosis by binding to the3’UTR of target mRNA to inhibit its transcription. miRNA can stably exist in the circulating blood, and its expression is consistent with the normal population. Studies have been shown that the miRNA in serum can express disease-specific changes when the body is in disorder. Recent studies have shown that miRNAs play an important role in bone metabolism, and has increasingly become a hot research. Osteoporosis is a systemic disease, which is characterized by a decrease in bone formation, and increased bone resorption, bone fragility, and prone to brittle fracture. As age increases, osteoporosis incidence is increasing. The purpose of this study was to identify the differentially expressed miRNAs in the osteoporosis patients. And to explore the target genes of differentially expressed bone metabolism associated miRNAs and their mutual cross talks with corresponding miRNAs and its role in the development of osteoporosis. The result would contribute to the miRNAs regulating mechanisms, and provide the theoretical guidance and experimental evidence for future gene therapy of osteoporosis.Methods:1, The blood samples of three cases of osteoporosis patients and three cases of age and sex-matched normal control group of were collected in this study, and extracted serum by centrifugation. Then the total RNA was isolated and tested with Exiqon miRNA Arrays after passing quality control. The slides scanned data of DISH patients and normal control group, osteoporosis and normal control group as well as osteoporosis and DISH group were analyses to explore the differentially expressed miRNAs. The differences criteria in screening expression of miRNA:up or down miRNA were Foldchange value>=2, P-value<=0.05.2. The difference miRNA sequence were analyzed by searching the miRbase database; the Targetscan and miRbase database were used for the target gene prediction; the relationship between differentially expressed miRNA and cell signaling pathways were predicted through http://Diana.cslab.ece.ntua.gr/pathways/; searching the target gene annotation information in Pumbmed and NCBI database. The possible target miRNA were selected based on the differentially expressed miRNA results and the current research results in the literature.Results:1. The differentially expressed miRNAs were selected based on the differences criteria after passing the quality control of samples. Compared with normal controls,5up-regulated and2down-regulated miRNA were found in osteoporosis group.2. The target differentially expressed miRNAs were selected according to the predicted target genes of each miRNA and associated cell signaling pathway as well as the existing research data in the literature.2up-regulated miRNA were selected in osteoporosis:hsa-miR-1321and hsa-miR-103a-3p, respectively.3. The related osteoporosis target genes involved in pathogenesis are:NOTCH3, DTX1, WNT16, BTRC, AXIN2and WNT3A; and the related cell signaling pathways were the Notch pathway and the Wnt pathway.Conclusion:1. This study compared the circulating miRNA of osteoporosis with healthy controls, and successfully found that differentially expressed miRNAs. Some of these miRNAs are associated with the regulation of bone metabolism.2. Using bioinformatics techniques to analyze differentially expressed miRNA, combined with the existing research results in the literature, we found the possible targate serum miRNA in osteoporosis patients. The specific regulatory mechanisms of differentially expressed miRNA in osteoporosis is needed to be further explored in subsequent studies.