ABSTRACT
Objective:To detect expression level of miR-410 in patients with systemic lupus erythematosus (SLE),and to expose the role of miR-410 and its target genes by bioinformatics methods.Methods:Expression level of miR-410 were detected by quantitative RT-PCR in peripheral blood mononuclear cells of SLE patients,and miR-410 sequence,its target genes and Genecards database were analyzed,and analysis of GO enrichment and KEGG Pathway was further performed.Results:miR-410 expression was significantly reduced in SLE patients,and its nucleotide sequence was highly conserved among species.These genes that were predicted to be regulated by miR-410 and associated with LE pathogenesis,included FASLG,CSF2,IFNAR2,MAPK1,PLCG2,IL4 and other genes.Analysis of GO enrichment revealed that miR-410's target genes were involved in cell growth,proliferation,programmed cell death,cell differentiation,immune system development and other biological activities.Analysis of KEGG Pathway showed that the target genes of miR-410 were significantly enriched in a series of signaling pathways including pathways in cancer,cytokine-cytokine receptor interaction,glioma,melanoma,TGF-β and JAK-STAT signaling pathway.Conclusion:miR-410 maybe directly regulate its target molecules,mediate various signal pathway networks,thus participate in the occurrence and development of SLE.
ABSTRACT
@#AIM: To study the effect of miR-410 on the regulation of angiotensin Ⅱ type 1 receptor(AT1R)in retinal pigment epithelium(RPE)cells of age-related macular degeneration(AMD)patients. <p>METHODS: The experiment was divided into AMD patients, cataract patients and normal people group. AT1R was the target gene of miR-410 by bioinformatics, and the normal RPE cells were cultured in the simulated microenvironment of AMD and cataracts and the expression of miR-410 was detected. Then miR-410 mimics was transfected into cells, and the expression of mRNA and protein of AT1R were detected by Q-PCR and Western blot respectively. The relationship between miR-410 and AT1R was confirmed by the dual luciferase reporter assay. <p>RESULTS: The miR-410 expression of in RPE cells with AMD was significantly reduced(<i>P</i>=0.0006, 0.0008)compared with cataract and normal controls. The miR-410 can regulate the function of AT1R by dual luciferase reporter gene experiment and the inhibition rate was about 40%. In addition, miR-410 inhibition rate was about 40%-50% to AT1R mRNA and protein expression by cell experiment.<p>CONCLUSION: AT1R was a target gene of miR-410 in cell experiments, and it is demonstrated that increasing the expression of miR-410 in RPE cells with AMD can suppress the expression of AT1R.
ABSTRACT
Retinal pigment epithelium (RPE) is a major component of the eye. This highly specialized cell type facilitates maintenance of the visual system. Because RPE loss induces an irreversible visual impairment, RPE generation techniques have recently been investigated as a potential therapeutic approach to RPE degeneration. The microRNA-based technique is a new strategy for producing RPE cells from adult stem cell sources. Previously, we identified that antisense microRNA-410 (anti-miR-410) induces RPE differentiation from amniotic epithelial stem cells. In this study, we investigated RPE differentiation from umbilical cord blood-derived mesenchymal stem cells (UCB-MSCs) via anti-miR-410 treatment. We identified miR-410 as a RPE-relevant microRNA in UCB-MSCs from among 21 putative human RPE-depleted microRNAs. Inhibition of miR-410 induces overexpression of immature and mature RPE-specific factors, including MITF, LRAT, RPE65, Bestrophin, and EMMPRIN. The RPE-induced cells were able to phagocytize microbeads. Results of our microRNA-based strategy demonstrated proof-of-principle for RPE differentiation in UCB-MSCs by using anti-miR-410 treatment without the use of additional factors or exogenous transduction.