圆锥角膜关键差异基因的鉴定与生物信息学分析

IDENTIFICATION AND BIOINFORMATICS ANALYSIS OF KEY DIFFERENTIALLY EXPRESSED GENES IN KERATOCONUS

目的鉴定圆锥角膜(KC)发病过程中的差异表达基因(DEGs),探讨KC潜在发病机制。方法分别收集5对KC与正常角膜捐献者的角膜组织,通过mRNA表达谱芯片筛选DEGs;借助KEGG通路富集分析明确其生物学功能;通过RT-qPCR验证候选DEGs的表达水平。结果共筛选出1885个DEGs,其功能与TGF-β信号通路、ErbB信号通路、鞘磷脂信号通路、谷氨酸能突触等有关。RT-qPCR验证显示,丝氨酸/苏氨酸激酶25(STK25)、酰基辅酶A结合结构域蛋白4(ACBD4)、锌指蛋白基因ZBTB38、组织蛋白酶B(CTSB)、3-羟基-3-甲基戊二酰辅酶A还原酶(HMGCR)、血小板凝血酶敏感蛋白的解聚蛋白样金属蛋白酶6(ADAMTS6)、DLG相关蛋白1(DLGAP1)等候选基因的表达水平与芯片结果一致。结论明确了KC差异基因表达谱,为后续KC发病机制的深入研究提供了基础数据。

Objective To identify the differentially expressed genes(DEGs)in the pathogenesis of keratoconus(KC)and explore the potential pathogenesis of KM.Methods Five pairs of corneal samples each from patients with KC and healthy donors were collected.mRNA microarray analysis was performed to select the DEGs,followed by KEGG pathway analysis to determine their biological functions.RT-qPCR was used to verify the expression of the candidate DEGs.Results A total of 1885 DEGs were detected.They were functionally associated with the TGF-βsignaling pathway,ErbB signaling pathway,sphingomyelin signaling pathway,and glutamatergic synapse.RT-qPCR revealed consistent results with the microarray analysis in the expression of the candidate DEGs including the serine/threonine protein kinase 25(STK25),acyl-CoA binding domain containing 4(ACBD4),zinc finger and BTB domain containing 38(ZBTB38),cathepsin B(CTSB),3-hydroxy-3-methylglutaryl-CoA reductase(HMGCR),ADAM metallopeptidase with thrombospondin type 1 motif 6(ADAMTS6),and DLG associated protein 1(DLGAP1).Conclusion We defined the expression profiles of DEGs in KC,providing basic data for subsequent in-depth investigation of the pathogenesis of KC.