[关键词]
[摘要]
目的:通过检测人翼状胬肉组织及正常结膜组织中miR-486-3p的表达情况,探寻其在翼状胬肉发生发展中可能的作用机制。
方法:于2018-09/2019-12收集原发性翼状胬肉患者69例69眼,收集术中切除的翼状胬肉组织和术眼正常结膜组织,采用RT-PCR定量检测标本中miR-486-3p的相对表达量,并利用 Targetscan数据库、miWalk3.0数据库、miRDB数据库对miR-486-3p进行靶基因预测后将得到的潜在靶基因取交集; 采用DAVID数据库对miR-486-3p潜在靶基因进行功能和通路富集分析; 采用String网站对miR-486-3p潜在靶基因进行互作分析。
结果:miR-486-3p在翼状胬肉组织中的相对表达量\〖(6.183±1.366)×10-6\〗与正常结膜组织的相对表达量\〖(7.930±1.394)×10-5\〗有显著差异(P<0.0001)。通过对其靶基因预测及生物信息学分析发现miR-486-3p潜在靶基因436个,其生物学功能主要富集于对RNA聚合酶Ⅱ启动子转录的调控、囊泡介导的转运和转录调控、DNA依赖性RNA代谢过程的调控。KEGG通路主要富集于神经轴突导向(Axon guidance)通路及溶酶体通路上,其中Axon guidance通路可能对翼状胬肉的发生发展起着重要的调控作用。PPI网络分析进一步表明Axon guidance通路中的关键基因ABL1、PLXNA1对翼状胬肉具有重要作用。
结论:miR-486-3p可能通过SLIT(神经导向因子Slit)/Robo(回旋引导受体)和SEMA3A(神经导向因子Semaphorin 3A)/PLXNA1(丛蛋白受体 A1)等Axon guidance通路致翼状胬肉的新生血管异常,从而参与翼状胬肉的发生、发展。
[Key word]
[Abstract]
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.
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.
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(P<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.
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.
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[基金项目]
国家自然科学基金项目(No.81770898); 武汉市卫生和计划生育委员会科研项目(No.WX18Q46)