Department of Pediatrics, First Affiliated Hospital of Guangxi Medical University, Guangxi Key Laboratory of Thalassemia Research, Nanning, Guangxi, China (mainland).
Guangxi Medical University, Nanning, Guangxi, China (mainland).
Med Sci Monit. 2019 Sep 21;25:7079-7086. doi: 10.12659/MSM.915946.
BACKGROUND Higher fetal hemoglobin (HbF) levels can ameliorate the clinical severity of ß-thalassemia. The use of integrative strategies to combine results from gene microarray expression profiling, experimental evidence, and bioinformatics helps reveal functional long noncoding RNAs (lncRNAs) in ß-thalassemia and HbF induction. MATERIAL AND METHODS In a previous study, a microarray profiling was performed of 7 individuals with high HbF levels and 7 normal individuals. Thirteen paired samples were used for validation. lncRNA NR_001589 and uc002fcj.1 were chosen for further research. The quantitative reverse transcription-PCR was used to detect the expression levels of 2 lncRNAs. The Spearman correlation test was employed. The nuclear and cytoplasmic distribution experiment in K562 cells was used to verify the subcellular localization of 2 lncRNAs. Potential relationships among lncRNAs, predicted microRNAs (miRNAs), and target gene HBG1/2 were based on competitive endogenous RNA theory and bioinformatics analysis. RESULTS Average expression levels of NR_001589 and uc002fcj.1 were significantly higher in the high-HbF group than in the control group. A positive correlation existed between NR_001589, uc002fcj.1, and HbF. The expression of NR_001589 was in both the cytoplasm and the nucleus, mostly (77%) in the cytoplasm. The expression of uc002fcj.1 was in both the cytoplasm and the nucleus; the cytoplasmic proportion was 43% of the total amount. A triple lncRNA-miRNA-mRNA network was established. CONCLUSIONS Novel candidate genetic factors associated with the HBG1/2 expression were identified. Further functional investigation of NR_001589 and uc002fcj.1 can help deepen the understanding of molecular mechanisms in ß-thalassemia.
较高的胎儿血红蛋白 (HbF) 水平可以改善 β-地中海贫血的临床严重程度。整合基因芯片表达谱、实验证据和生物信息学的综合策略有助于揭示 β-地中海贫血和 HbF 诱导中的功能性长非编码 RNA (lncRNA)。
在之前的一项研究中，对 7 名 HbF 水平较高的个体和 7 名正常个体进行了基因芯片表达谱分析。使用 13 对样本进行验证。选择 lncRNA NR_001589 和 uc002fcj.1 进行进一步研究。采用实时定量逆转录-PCR 检测 2 个 lncRNA 的表达水平。采用 Spearman 相关检验。在 K562 细胞中进行核质分布实验，验证 2 个 lncRNA 的亚细胞定位。根据竞争内源性 RNA 理论和生物信息学分析，预测 lncRNA 与预测 microRNA (miRNA) 和靶基因 HBG1/2 之间的潜在关系。
NR_001589 和 uc002fcj.1 的平均表达水平在高-HbF 组显著高于对照组。NR_001589、uc002fcj.1 与 HbF 呈正相关。NR_001589 表达于细胞质和细胞核中，主要位于细胞质中（77%）。uc002fcj.1 表达于细胞质和细胞核中，细胞质部分占总量的 43%。建立了一个三 lncRNA-miRNA-mRNA 网络。
鉴定出与 HBG1/2 表达相关的新型候选遗传因子。进一步研究 NR_001589 和 uc002fcj.1 的功能有助于加深对 β-地中海贫血分子机制的理解。
