High Glucose And/or Lipid Levels Down-Regulate MiR-145 That Targets Osteoprotegerin And Modulates Cell Apoptosis, Proliferation And Chromosome Missegregation

Partâ… High glucose and/or lipid levels dysregulate the expression in both cytokines and miRNAs in monocytes/macrophages.OBJECTIVE:To explore the differential expressions of cytokines and microRNAs of human monocytes/macrophages under high glucose and/or lipid conditions.METHODS:We performed miRNA microarrays and cytokine antibody arrays under high glucose and/or lipid intervention in monocytic THP-1 cells.We confirmed the antibody arrays by using ELISA and western blotting.The expression levels of miRNAs were confirmed with real-time PCR.RESULTS:High glucose and/or lipid induced 108 cytokines dysregulation. IL-12p40 and OPG were up-regulated by using ELISA and western blotting,respectively.On the other hand,real-time PCR showed that miR-128a and miR-145 were down-regulated.CONCLUSION:High glucose and/or lipid can dysregulate the expression in both cytokines and miRNAs in monocytes/macrophages. Partâ…¡miR-145 targets osteoprotegerinOBJECTIVE:To clarify the role ofmiRNAs in the regulation of cytokine expressions.METHODS:Serum OPG levels were determined by ELISA,the relative expression of miR-145 was evaluated with real-time PCR. Transfection of miR-145 inhibitor or precursor was employed with elecroporation or lipofectamine method.Western blotting or immunofluorescence analysis was used to assess the expressions of OPG.The interaction between miR-145 and OPG 3'UTR was evaluated by luceferase assay system.RESULTS:Serum OPG levels were negatively correlated with the amounts of miR- 145 in peripheral blood mononuclear cells. Western blotting and immunofluorescence analysis revealed that OPG was up-regulated after transfecting miR-145 inhibitor,by contrast,over-expression of miR-145 reduced OPG levels,miR-145 directly bound OPG 3'UTRCONCLUSION:miR-145 targets osteoprotegerin expression. Partâ…¢miR-145 modulates cell proliferation,apoptosis and chromosome segregation.OBJECTIVE:To evaluate the role of miR-145 in cell proliferation, apoptosis and chromosome segregation.METHODS:Cell proliferation was assessed by XTT and growth curve. For apoptosis analyses,we performed western blotting and TUNEL assay to verify the proapoptotic role of miR-145. Chromosome segregation was analyzed with immunofluorescence.RESULTS:Inhibition of miR-145 promoted cell proliferation and impeded apoptosis.Over-expression of miR-145 signaled apoptosis independently of p53 and resulted in widespread chromosome missegregation.CONCLUSION:miR-145 modulates cell proliferation,apoptosis and chromosome segregation.