有吸烟史的口腔白斑病患者microRNA异常表达及癌变相关生物信息学分析

Microarray combined with bioinformatics analysis identifies the aberrantly expressed microRNA related to cancerization in oral leukoplakia patients with a history of tobacco smoking

目的研究有吸烟史的口腔白斑病(oral leukoplakia,OLK)患者病损组织microRNA(miRNA)的异常表达,并对差异miRNAs进行癌变相关生物信息学分析。方法利用人miRNA OneArray?v5.1微阵列芯片检测吸烟组(n=6)和非吸烟组(n=4)OLK石蜡标本间差异表达的miRNA,并运用生物信息学方法从中筛选出与OLK癌变密切相关的目标miRNA,随后对目标miRNA的靶基因进行基因本体数据库(gene ontology,GO)富集分析和KEGG通路分析以预测其可能的作用机制。结果吸烟组与非吸烟组OLK组织中共检出174个差异表达的miRNAs(P<0.05),其中,39个miRNAs在吸烟组表达上调(log2(FC)≥0.585),135个miRNAs表达下调(log2(FC)≤-0.585)。运用生物信息学方法从中筛选出8个与OLK癌变密切相关的目标miRNAs,包括:hsa-miR-6857-5p、hsa-miR-6745、hsa-miR-15a-5p、hsa-miR-363-5p、hsa-miR-6867-3p、hsa-miR-95-3p、hsa-miR-3607-5p、hsa-miR-9500。对上述目标miRNAs的靶基因行GO富集分析,结果显示大部分基因参与调控细胞增殖、凋亡、刺激反应等生物学活动。KEGG通路分析发现上述基因高度相关的作用通路为癌信号通路、PI3K-Akt及FoxO信号通路。结论吸烟可通过改变OLK组织miRNA表达水平进而改变特定基因功能促进白斑癌变进程。

Objective To compare the aberrantly expressed miRNA in OLK tissue from patients with smoking habit and non-smoking control subjects by microarray chip,and perform bioinformatics analyses of these differential miRNAs related to OLK cancerization.Methods Human miRNA OneArrayv5.1 microarray chip was used to detect and screen the aberrantly expressed miRNAs in OLK paraffin specimens between smoking and non-smoking groups.The target miRNAs closely related to OLK cancerization were further screened by bioinformatics analyses.Then the target genes of these targeted miRNAs were subjected to GO enrichment analysis and KEGG pathway analysis to explore the possible mechanism of target miRNAs in OLK cancerization.Results A total of 174 differentially expressed miRNAs were detected,of which 39 miRNAs were up-regulated in the smoking group(log2(FC)≥0.585),and 135 miRNAs were down-regulated(log2(FC)≤-0.585).Among them,8 miRNAs related to OLK cancerization were screened out,including hsa-miR-6857-5p,hsa-miR-6745,hsa-miR-15a-5p,hsa-miR-363-5p,hsa-miR-6867-3p,hsa-miR-95-3p,hsa-miR-3607-5p and hsa-miR-9500.Gene-ontology(GO)enrichment analysis of genes targeted by the above 8 differential miRNAs found that most of the target genes were involved in cell proliferation,apoptosis,and stimulation response processes.Kyoto Encyclopedia of Genes and Genomes(KEGG)enrichment pathway analysis showed signal pathways highly related to these target genes were pathways in cancer,PI3K-Akt and FoxO pathways.Conclusions Smoking can change the function of specific genes by altering some miRNA expression levels in OLK tissue,thus participating in OLK carcinogenesis.