Hematology Laboratory, Shengjing Hospital of China Medical University, Shenyang, China.
Int J Lab Hematol. 2020 Dec;42(6):858-865. doi: 10.1111/ijlh.13305. Epub 2020 Oct 2.
Long noncoding RNAs (lncRNAs) play significant roles in the carcinogenesis and progression of human tumors. The aim of this work was to evaluate the expression level of lncRNAs in adult acute myeloid leukemia (AML) patients and to explore the function of candidate lncRNA PCAT18 in clinical diagnosis and prognosis of AML.
Noncoding RNA screening was performed by second-generation sequencing on adult newly diagnosed AML patients and healthy controls. GO and KEGG pathway analysis were performed for functional analysis of differentially regulated lncRNAs. The expression of candidate lncRNA PCAT18 was detected by real-time PCR. In K-562 and THP-1 cells, proliferation, and cell cycle analysis were performed using CCK-8 assay and Cell Cycle Assay Kit.
The lncRNA expression profile of AML patients and healthy controls showed that six upregulated (>5-fold) and eight downregulated (<0.2-fold) lncRNAs. The candidate lncRNA PCAT18 showed higher expression in AML patients with NPM1 mutation and favorable-risk. RT-PCR revealed a significant increase of PCAT18 expression comparing to control cells. PCAT18 overexpression obviously promoted cell proliferation and PCAT18 knockdown decreased cell proliferation. The fraction of the S phases was increased in the PCAT18 overexpression group and decreased in the PCAT18 knockdown group.
The candidate lncRNA PCAT18 in our study was firstly found upregulated in AML samples, that its overexpression promoted cell proliferation and G1/S transition. Further study of PCAT18 and its target mRNAs are needed to confirm the mechanism of PCAT18 in AML. PCAT18 may act as diagnostic and prognosis biomarker for AML.
长链非编码 RNA（lncRNAs）在人类肿瘤的发生和发展中发挥重要作用。本研究旨在评估 lncRNAs 在成人急性髓系白血病（AML）患者中的表达水平，并探讨候选 lncRNA PCAT18 在 AML 临床诊断和预后中的作用。
采用第二代测序技术对新诊断的成人 AML 患者和健康对照进行非编码 RNA 筛选。对差异表达的 lncRNAs 进行 GO 和 KEGG 通路分析，以进行功能分析。采用实时 PCR 检测候选 lncRNA PCAT18 的表达。在 K-562 和 THP-1 细胞中，通过 CCK-8 法和细胞周期试剂盒检测细胞增殖和细胞周期。
AML 患者和健康对照的 lncRNA 表达谱显示，有 6 个上调（>5 倍）和 8 个下调（<0.2 倍）的 lncRNAs。候选 lncRNA PCAT18 在 NPM1 突变和低危 AML 患者中表达较高。与对照细胞相比，RT-PCR 显示 PCAT18 表达明显增加。PCAT18 过表达明显促进细胞增殖，而 PCAT18 敲低则降低细胞增殖。PCAT18 过表达组 S 期细胞比例增加，PCAT18 敲低组 S 期细胞比例减少。
本研究首次发现候选 lncRNA PCAT18 在 AML 样本中上调，其过表达促进细胞增殖和 G1/S 期转换。需要进一步研究 PCAT18 及其靶 mRNAs，以确认 PCAT18 在 AML 中的作用机制。PCAT18 可能作为 AML 的诊断和预后生物标志物。
