通过 RNA 测序进行全面的长非编码 RNA 表达谱分析揭示了急性髓细胞白血病风险的潜在生物标志物。

Comprehensive long non-coding RNA expression profiling by RNA sequencing reveals potential biomarkers for acute myeloid leukemia risk.

出版信息

Cancer Biomark. 2019;26(1):93-108. doi: 10.3233/CBM-190215.

DOI:10.3233/CBM-190215

Abstract

OBJECTIVE

The aim of this work was to extensively explore the long non-coding RNA (lncRNA) expression profiles in acute myeloid leukemia (AML) and to propose candidate lncRNAs with predictive value for AML risk.

METHODS

The bone marrow mononuclear cell samples from 10 AML patients and 10 age and gender matched controls were obtained and subjected to next-generation RNA sequencing. Then, the top 5 upregulated and top 5 downregulated lncRNAs in AML patients compared with controls were selected for further quantitative polymerase chain reaction (qPCR) validation in 40 AML patients and 40 age and gender matched controls. The effect of candidate lncRNA RP11-342M1.7 on proliferation and apoptosis in AML cells was further investigated.

RESULTS

RNA sequencing observed 216 upregulated and 412 downregulated lncRNAs in AML patients compared with controls. Enrichment analyses exhibited that these differentially expressed lncRNAs were involved in neoplastic signaling pathways such as cAMP and MAPK signaling pathways. In further qPCR validation, lncRNA RP11-342M1.7 and lncRNA CDCA4P3 were upregulated, while lncRNA CES1P1, lncRNA AC008753.6 and lncRNA RP11-573G6.10 were downregulated in AML patients compared with controls. Multivariate logistic regression analysis disclosed that lncRNA RP11-342M1.7, lncRNA CES1P1 and lncRNA AC008753.6 were independent predictive factors for AML risk, and most importantly, the combination of these three lncRNAs was of remarkably good predictive value for AML risk (AUC: 0.901; 95% CI: 0.835-0.966). Besides, lncRNA RP11-342M1.7 was correlated with higher CR while lncRNA AC008753.6 and lncRNA CTD-2562J15.6 were correlated with lower CR. LncRNA RP11-342M1.7 knockdown suppressed cell proliferation by promoting cell apoptosis in AML cells.

CONCLUSIONS

This study reveals the comprehensive lncRNAs expression profiles in AML, and proposes candidate lncRNAs that are potential biomarkers for AML risk.

摘要

目的

本研究旨在广泛探索急性髓系白血病（AML）中长链非编码 RNA（lncRNA）的表达谱，并提出具有预测 AML 风险的候选 lncRNA。

方法

采集 10 例 AML 患者和 10 例年龄和性别匹配的对照者的骨髓单个核细胞样本，进行下一代 RNA 测序。然后，选择 AML 患者与对照组相比上调幅度最大的前 5 个和下调幅度最大的前 5 个 lncRNA 进行进一步的定量聚合酶链反应（qPCR）验证，验证对象为 40 例 AML 患者和 40 例年龄和性别匹配的对照者。进一步研究候选 lncRNA RP11-342M1.7 对 AML 细胞增殖和凋亡的影响。

结果

与对照组相比，RNA 测序观察到 AML 患者中有 216 个上调和 412 个下调的 lncRNA。富集分析表明，这些差异表达的 lncRNA 参与了致癌信号通路，如 cAMP 和 MAPK 信号通路。在进一步的 qPCR 验证中，与对照组相比，lncRNA RP11-342M1.7 和 lncRNA CDCA4P3 上调，而 lncRNA CES1P1、lncRNA AC008753.6 和 lncRNA RP11-573G6.10 下调。多变量逻辑回归分析显示，lncRNA RP11-342M1.7、lncRNA CES1P1 和 lncRNA AC008753.6 是 AML 风险的独立预测因素，更重要的是，这三个 lncRNA 的组合对 AML 风险具有极好的预测价值（AUC：0.901；95%CI：0.835-0.966）。此外，lncRNA RP11-342M1.7 与较高的完全缓解率相关，而 lncRNA AC008753.6 和 lncRNA CTD-2562J15.6 与较低的完全缓解率相关。lncRNA RP11-342M1.7 的敲低通过促进 AML 细胞凋亡来抑制细胞增殖。