Wnt/β-catenin信号通路相关长链非编码RNA在肺结核患者外周血中的表达及其意义

Expression and significance of Wnt/β-catenin signaling pathway-related long non-coding RNA in peripheral blood of patients with pulmonary tuberculosis

目的 探讨Wnt/β-catenin信号通路相关长链非编码RNA（lncRNA）在肺结核患者外周血单核细胞（PBMCS）中的表达及其临床意义。 方法 选取初治结核患者、结核杆菌感染者、未感染结核杆菌者各50例，采集其外周血、分离单核细胞、提取RNA，采用mRNA芯片检测，通过聚类分析、GO和KEGG pathway分析，筛选肺结核患者差异表达的Wnt/β-catenin信号途径相关lncRNA。以实时荧光定量PCR（qRT-PCR）技术检测相关lncRNA表达的变化情况和各组样本中的Wnt/β-catenin信号通路活化情况。 结果 mRNA芯片共检出差异表达的lncRNA 4 487个：其中上调2 756个，下调1 731个。通过GO 和KEGG pathway 分析筛选Wnt/β-catenin信号通路途径相关差异表达的lncRNA FR255266、NONHSAT006952、TCONS_l2_00020446做进一步分析。REAL TIME PCR 检测结果显示，与正常对照组相比，结核杆菌潜伏感染者外周血 PBMCS 中lncRNA FR255266、 TCONS_l2_00020446 表达显著升高, 且在肺结核患者中进一步上调，而lncRNA NONHSAT006952 在肺结核患者中显著降低；结核杆菌潜伏感染者和肺结核患者Wnt/β-catenin信号通路相关mRNA：TCF, GSK-3β和β-catenin的表达显著上调，以上差异均有统计学意义（ P 值均＜0.05）。 结论 结核菌感染后，机体内 Wnt/β-catenin 信号通路途径相关的一系列lncRNA表达发生显著变化，须进一步探究其在结核病中的生物学功能及其分子机制，可将其作为结核病的生物预警、诊断及精准治疗提供新的靶点。

Objective To investigate the expression and clinical significance of long non-coding RNA （lncRNA） related to Wnt/β-catenin signaling pathway in peripheral blood mononuclear cells （PBMCs） of patients with pulmonary tuberculosis（TB）. Methods A total of 50 patients of initial treatment of pulmonary tuberculosis, 50 cases with tuberculosis bacilli infection and 50 cases without tuberculosis bacilli infection were selected. Peripheral blood were collected, monocytes were isolated, RNA was extracted and subjected to mRNA chip assay. The differential expression of Wnt/β-catenin signal pathway-related lncRNAs among tuberculosis patients were screened by cluster analysis, GO and KEGG pathway analysis. Real-time quantitative PCR （qRT-PCR） was used to detect the expression of related lncRNA and the activation of Wnt/β-catenin signaling pathway in each group. Results A total of 4 487 lncRNA expressed differentially on mRNA chip, among which 2 756 were up regulated and 1 731 were down regulated. The differentially expressed lncRNA FR255266, NONHSAT006952 and TCONS_l2_00020446 related to Wnt/β-catenin signaling pathway were screened by GO and KEGG pathway analysis for further analysis. qRT-PCR showed that the expression of lncRNA FR255266 and TCONS_l2_00020446 in PBMCs of latent infection tuberculosis cases were significantly higher than that of normal control group and further increased among tuberculosis patients; while the expression of lncRNA NONHSAT006952 was significantly lower among tuberculosis patients. The expressions of Wnt/β-catenin signaling pathway-related genes such as TCF, GSK-3βandβ-catenin were significantly increased among tuberculosis patients, all differences were statistically significant （all P 〈0.05）. Conclusion The expression of a series of lncRNA related to Wnt/β-catenin signaling pathway changes significantly upon tuberculosis infection. Further exploration of its biological function and molecular mechanism in tuberculosis will provide a new target for biological warning, diagnosis and precise treatment of tuberculosis.