Expression of miR-486-3p and bioinformatics analysis of its potential target genes in human pterygium / 国际眼科杂志(Guoji Yanke Zazhi)

@#AIM: To detect the expression of miR-486-3p in human pterygium tissue and normal conjunctival tissue and explore the possible mechanism of miR-486-3p in the development of pterygium. <p>METHODS: Totally 69 patients 69 eyes with primary pterygium treat in Zhongnan Hospital of Wuhan University and Hankou Aier Eye Hospital from September 2018 to December 2019 were collected by excision of pterygium during surgery(experimental group). At the same time, a total of 69 patients with normal conjunctival tissue of their same eyes were taken as control group during surgery. The relative expression levels of miR-486-3p in the experimental group and the control group were quantitatively detected by RT-PCR. The Targetscan database, miWalk3.0 database and miRDB database were used to predict the potential target genes of miR-486-3p. DAVID database was used to analyse and enrich the function and pathway of the potential target genes of miR-486-3p. The String website performed an interactional analysis of the potential target genes of miR-486-3p. <p>RESULTS: The relative expression level of miR-486-3p in the experimental group(6.183±1.366)×10^-6 was significantly different from that in the control group(7.930±1.394)×10^-5(<i>P</i><0.0001). By the prediction of their target genes and bioinformatical analysis, a total of 436 potential target genes of miR-486-3p were found. The biological functions were mainly concentrated in the regulation of RNA polymerase II promoter transcription, vesicle-mediated transport, transcriptional regulation and the regulation of DNA-dependent RNA metabolism. The Kyoto Encyclopedia of Genes and Genomes(KEGG)pathway was mainly enriched in the Axon guidance pathway and lysosomal pathway. And the Axon guidance pathway might play an important regulatory role in the occurrence and development of pterygium. PPI network analysis further elucidated that the key genes of ABL1 and PLXNA1(cell protein receptor A1)play an important role in the Axon guidance pathway for pterygium. <p>CONCLUSION: MiR-486-3p might be involved in the occurrence and development of pterygium through SLIT(neuro-targeting factor)/Robo(rotatory guide receptor)and SEMA3A(neuro-guiding factor Semaphorin 3A)/ PLXNA1 of Axon guidance pathways, which resulted in the abnormal new blood vessels of pterygium.

Expression of miR-486-3p in psoriatic lesions and its effect on keratin 17 expression / 中华皮肤科杂志

Objective To determine the expression of miR-486-3p in psoriatic lesions and healthy human skin and to estimate its effect on keratin 17 (K17) expression in HaCaT human keratinocytes.Methods Bioinformatics was used to predict microRNAs that may affect the expression of K17.Tissue samples were obtained from the lesions of 10 patients with psoriasis and normal skin of 10 healthy human controls.RNA was extracted from these tissue samples and reversely transcribed into cDNA with the addition of a Poly (A) tail.Then,real time quantitative PCR was performed to measure the expression of miR-486-3p.Cultured keratinocytes were transfected with miR-486-3p mimics or negative control,and Western blot was performed to determine K17 expression at 48 hours after the transfection.To evaluate the inhibitory effect of miR-486-3p on K17 expression,cultured 293T cells were transfected with the plasmid containing K17 3' untranslated region (UTR) seed sequence,three plasmids containing the complete deletion,interval mutation or double repeats of the seed sequence,or negative control plasmid.At 24 hours after the transfection,a dualluciferase reporter (DLR) assay was performed to quantify the expression of K17.Results Real time PCR showed that the expression level of miR-486-3p was significantly lower in psoriatic lesions than in the normal skin (0.211 ± 0.120vs.0.555 ± 0.425,t =2.62,v =9,P ＜ 0.05).The HaCaT cells transfected with the mimics of miR-486-3p exhibited decreased expression of K17 compared with those transfected with the negative control.DLR assay revealed that the expression level (fluorescence intensity) of K17 in the negative control group was significantly higher than that in the 293T cells transfected with the seed sequence and those with the double repeats of the seed sequence (100.00％ vs.65.31％ ± 6.32％ and 54.18％ ± 10.01％ respectively,both P ＜ 0.05),but did not differ from that in the 293T cells transfected with the complete deletion and interval mutation of the seed sequence (100.00％ vs.114.77％ ± 16.14％ and 110.21％ ± 12.99％ respectively,both P ＞ 0.05).Conclusions The expression of miR-486-3p,which may inhibit K17 expression by binding to the seed sequence of K17 3'UTR,is lower in psoriatic lesions than in normal skin.The decreased expression and inhibitory effect of miR-486-3p may be implicated in the initiation and progression of psoriasis.