结核性胸膜炎患者胸膜组织中环状RNA表达谱初步分析
Analysis of circular RNA expression profile in pleural tissues of patients with tuberculous pleurisy
目的:分析结核性胸膜炎患者胸膜组织中环状RNA(circRNA)的表达谱,并预测其靶基因,初步探讨circRNA与结核性胸膜炎的关系。方法:本研究纳入了结核性胸膜炎患者和对照组各13例,所有患者均通过胸腔镜进行胸膜活检。应用高通量测序技术对结核性胸膜炎组织与正常胸膜组织各3例进行circRNA检测,用edgeR对基因进行了差异表达分析,剩余标本进行实时荧光定量聚合酶链反应(qRT-PCR)验证,用miRanda软件预测差异表达circRNA相关的微小RNA(miRNA)。结果:高通量测序结果显示,结核性胸膜炎组织中存在差异表达的circRNA有134个,其中53个表达上调,81个表达下调。挑选6个有显著差异的circRNA进行qRT-PCR验证,结果显示hsa_circ_0008433在结核性胸膜炎组织中的表达上调( P<0.05),hsa_circ_0008234、hsa_circ_0001368在结核性胸膜炎组织中的表达下调( P<0.05),结果与高通量测序结果一致。其余3个circRNA的表达水平差异无统计学意义。miRanda软件预测hsa-miR-17、hsa-miR-20b-3p等是hsa_circ_0008234的靶基因,其可能在结核性胸膜炎的发生发展中起调控作用。 结论:结核性胸膜炎患者胸膜组织中circRNA表达谱发生了变化,这些差异表达的circRNA可能与结核性胸膜炎的发生发展密切相关。
更多Objective:To explore the expression profile of circular RNA (circRNA) in pleural tissues of patients with tuberculous pleurisy and the relationship between circRNA and tuberculous pleurisy.Methods:13 patients with tuberculous pleurisy and 13 patients in the control group were included.All patients underwent thoracoscopic pleural biopsy.CircRNA was detected in three cases of tuberculous pleurisy and three cases of normal pleural tissue by high-throughput sequencing, the differential expression of circRNA was analyzed with edgeR.Quantitative real-time polymerase chain reaction (qRT-PCR) was carried out for the remaining samples to verify the results.miRanda was used to predict microRNA related to differentially expressed circRNA.Results:A list of 134 circRNAs was differentially expressed among tuberculous pleurisy, 53 up-regulated and 81 down-regulated.Six circRNAs with significant differences were selected for qRT-PCR validation.It was shown that hsa_circ_0008433 was up-regulated in pleura infected with tuberculosis ( P<0.05), while hsa_circ_0008234 and hsa_circ_0001368 were down-regulated ( P<0.05), which was consistent with the high-throughput sequencing result.It was shown that hsa-miR-17 and hsa-miR-20b-3p might be target genes of hsa_circ_0008234, which might play a regulatory role in the occurrence and development of tuberculous pleurisy. Conclusions:The expression profile of circRNA in tuberculous pleurisy is changed.These differentially expressed circRNAs may be closely related to the occurrence and development of tuberculous pleurisy.
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