子痫前期胎盘组织环状RNA表达谱的鉴定、验证与分析

Identification and analysis profiles of circular RNAs in preeclamptic placenta

目的:描绘人胎盘组织中环状RNA(circular RNA,circRNA)的表达谱,发现并验证子痫前期胎盘组织异常表达的circRNA,研究分析其可能的调控网络。方法:利用高通量测序技术对子痫前期和正常胎盘组织中RNA进行测序和鉴定分析,并利用DeSeq2进行差异性分析。然后利用qRT-PCR在组织水平上对分析结果进行验证。随后利用生物信息学对验证的差异性circRNA进行GO分析和KEGG通路分析。结果:测序结果显示,与正常胎盘组织相比,重度子痫前期胎盘共检测出300种异常表达的circRNA,包括151种高表达和149种低表达。qRT-PCR验证结果显示,hg38circ0014736 (1.418 0±0.106 5 vs. 1.044 0±0.127 1,P=0.035)和hsacirc0015382 (0.895 8±0.180 2 vs. 0.688 8±0.025 3,P=0.025)在子痫前期胎盘组织中高表达,hsacirc0007121(0.395 8±0.200 4 vs. 1.301 0±0.240 7,P=0.000)在子痫前期胎盘组织中明显下调。生物信息学分析显示这些异常表达的circRNA与预测到的靶miRNA及靶基因可能参与调节细胞增殖、缺氧应激、转录后调控等功能。结论:与正常胎盘组织相比,子痫前期胎盘组织存在异常表达的circRNA。生物信息学分析显示,这些异常表达的circRNA或可以通过某些通路参与子痫前期发病。

Objective:To describe the expression profile of circular RNA(circRNA)in human placental tissues,explore and identify abnormal circRNAs in preeclampsia(PE)placental tissue and study its possible regulating network.Methods:RNA sequencing and e-valuation analysis in PE placental tissue and normal placental tissue were performed by high-throughput RNA sequencing technology,DeSeq2 was used to analyze difference,and qRT-PCR was used to identify the analysis tissue.Bioinformatics was used to perform the gene ontology(GO)analysis and the Kyoto encyclopedia of genes and genomes(KEGG)pathway analysis in the identified differential circRNA.Results:According to the sequencing results,300 differentially expressed circRNAs were found in the severe PE placenta when compared with the normal placenta,including 151 kinds of upregulated circRNAs and 149 kinds of downregulatedcircRNAs.The qRT-PCR results showed that expression levels of hg38_circ_0014736(1.4180±0.1065 vs.1.0440±0.1271,P=0.035)and hsa_circ_0015382(0.8958±0.1802 vs.0.6888±0.0253,P=0.025)were significantly higher in the PE placenta tissue;expres-sion levels of hsa_circ_0007121(0.3958±0.2004 vs.1.3010±0.2407,P=0.000)were significantly decreased in PE placenta com-pared with normal placenta.Bioinformatics analysis revealed that those three circRNAs,predicted target miRNAs and target gene might have a relationship with cell proliferation,response to hypoxia and post-transcriptional regulation.Conclusion:Com-pared with the normal placental tissue,abnormal expressive circRNA exists in the PE placental tissue.According to the bioinformatics,those abnormal circRNAs may play a potential role in the pathogenesis of PE.