环状RNA在小鼠肺发育中的连续表达及功能预测

Continuous expression and functional prediction of circular RNA in mouse lung development

目的探讨环状RNA(circRNA)circ4:150439343|150477468和circ15:73330849|73343359在小鼠肺发育中的连续表达及潜在功能。方法根据肺发育分期,取孕16.5 d(E16.5 d)、孕18.5 d(E18.5 d)及生后2 d(P2 d)胎鼠及新生小鼠的肺组织,利用苏木精-伊红染色观察肺组织形态学;利用qRT-PCR技术检测晚期肺发育过程中circ4:150439343|150477468和circ15:73330849|73343359的表达情况;应用miRanda和TargetScan软件预测circRNA的靶向miRNA,然后对靶基因进行GO和KEGG分析并预测相应circRNA功能。结果在E16.5 d小鼠肺组织切片中观察到Ⅱ型肺泡上皮细胞,且逐渐增多,P2 d时肺泡迅速扩张,间质变薄,肺泡结构逐渐成熟;qRT-PCR结果显示circ4:150439343|150477468相对表达量随时间推移持续上调(P<0.05),circ15:73330849|73343359的相对表达量先下调再上调(P<0.05);宿主基因功能预测显示circRNA可参与Notch、PI3K-Akt和NF-κB等信号通路。结论circ4:150439343|150477468和circ15:73330849|73343359可通过Notch等信号通路参与肺发育的调控。

Objective To study the continuous expression and potential function of circular RNA(circRNA),circ4:150439343|150477468 and circ15:73330849|73343359,in mouse lung development.Methods According to the stage of lung development,lung tissue samples were collected from mice on embryonic day 16.5(E16.5),embryonic day 18.5(E18.5),and postnatal day 2(P2).Hematoxylin and eosin staining was performed to observe the morphology of lung tissue.Quantitative real-time PCR(qRT-PCR)was used to measure the mRNA expression of circ4:150439343|150477468 and circ15:73330849|73343359 during late lung development;miRanda and TargetScan were used to predict the target miRNAs of circRNAs,and then GO and KEGG analysis was performed for the target genes to predict the potential function of circRNAs.Results Type II alveolar epithelial cells were observed in the lung slices of E16.5 mice,with a gradual increase in number.On P2,the pulmonary alveoli expanded rapidly,the pulmonary interstitium became thinner,and the alveolar structure gradually became mature.The results of qRT-PCR showed that the relative expression of circ4:150439343|150477468 was continuously upregulated over time and the relative expression of circ15:73330849|73343359 was first downregulated and then upregulated(P<0.05).The KEGG and GO analysis showed that circRNAs were involved in the Notch,PI3K-Akt,and NF-κB signaling pathways.Conclusions Circ4:150439343|150477468 and circ15:73330849|73343359 can participate in lung development through the Notch signaling pathway.