Objective By detecting the expression of E26 transformation specific sequence 1(ETS1)gene and the genotype of its single nucleotide polymorphism(SNP)site rs4937333 in patients with osteoporosis and healthy individuals,this study aims to explore the relationship between the rs4937333 site on the ETS1 gene and its upstream regulatory factor miR-760 in the pathogenesis of osteoporosis.Methods Peripheral blood samples were collected from osteoporosis patients and healthy controls,followed by isolation of peripheral blood mononuclear cells(PBMCs),CD4+T cells,and CD19+B cells.The expression of ETS1 mRNA in different cell types was assessed using qPCR.Simultaneously,the genotype and allele frequency of the ETS1 gene polymorphism rs4937333 were determined using SNaPShot technology in a cohort consisting of 200 osteoporosis patients and 45 healthy controls.Additionally,recombinant psi-CHECK-2-ETS1-3'UTR WT and psi-CHECK-2-ETS1-3'UTR MT plasmids were constructed to evaluate firefly luciferase activity through a dual luciferase assay in cells.Furthermore,an ETS1 overexpression vector was generated and transfected into B lymphocytes to examine the expression levels of ETS1 using qPCR and Western blotting analysis.Results Compared to those in the healthy group,the expression levels of ETS1 in PBMCs,CD4+T cells,and CD19+B cells from osteoporosis patients was significantly reduced(all P＜0.01).Additionally,the proportion of C/T genotype at SNP locus rs4937333 in the 3'UTR region of the ETS1 gene was higher in patients than in healthy individuals.qPCR result demonstrated that patients with a C/T genotype had significantly lower ETS1 expression levels than in those with a C/C genotype(all P＜0.01).Luciferase assays revealed a significant decrease in luciferase activity when miR-760 mimic and wild-type 3'UTR of ETS1 were co-transfected(P＜0.01).qPCR and Western blotting experiments detected decreased expression levels of ETS1 during plasmid infection of B lymphocytes in mutant strains compared to wild-type strains(P＜0.01).Conclusion MiR-760 targets and regulates the rs4937333 site on the 3'UTR sequence of the ETS1 gene,reducing the expression of ETS1 and participating in the pathological occurrence of osteoporosis.
维普
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