Department of Hematology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, People's Republic of China.
Department of Hematology, The Second Affiliated Hospital of Nanchang University, Nanchang, Jiangxi Province, People's Republic of China.
Pathology. 2021 Aug;53(5):613-622. doi: 10.1016/j.pathol.2020.11.003. Epub 2021 Feb 6.
LncRNA PVT1 has been demonstrated to be upregulated in acute myeloid leukaemia (AML) patients and indicates a poor prognosis. Nevertheless, its role in AML remains obscure. This study investigated the regulatory role and potential mechanisms of PVT1 in the progression of AML. Expression of PVT1, miR-29 family and WAVE1 was detected by quantitative real-time polymerase chain reaction. CCK8 and EdU assays were performed to assess the proliferation of AML cells. Cell cycle and apoptosis were determined by propidium iodide (PI) staining and Annexin V/PI staining on a flow cytometer. Transwell assay was carried out to evaluate the migration and invasion abilities. The interaction between miR-29 family and PVT1/WAVE1 was confirmed by dual luciferase reporter assay and RNA immunoprecipitation assay. The protein levels of WAVE1, Bcl-2, Bax, cleaved Caspase 3, cyclin D1, and p21 were detected by western blotting. Xenograft transplantation was performed to determine the tumourigenicity of AML cell in vivo. PVT1 expression was significantly increased in AML patient samples and cells, which positively correlated with WAVE1 expression. Silencing of PVT1 restrained growth, migration and invasion, while inducing apoptosis of AML cells. Moreover, PVT1 acted as a sponge for miR-29 family to increase WAVE1 expression in AML cells. Overexpression of WAVE1 partly counteracted PVT1 knockdown-induced anti-tumour effects on AML cells in vitro and xenograft tumour in vivo. PVT1 facilitated the progression of AML via regulating miR-29 family/WAVE1 axis, which supported the conclusion that PVT1 may be a promising therapeutic target for AML.
长链非编码 RNA PVT1 在急性髓系白血病（AML）患者中呈上调表达，提示预后不良。然而，其在 AML 中的作用仍不清楚。本研究旨在探讨 PVT1 在 AML 进展中的调控作用及其潜在机制。采用实时定量聚合酶链反应检测 PVT1、miR-29 家族和 WAVE1 的表达。通过 CCK8 和 EdU 检测评估 AML 细胞的增殖。通过碘化丙啶（PI）染色和流式细胞术 Annexin V/PI 染色检测细胞周期和凋亡。Transwell 检测评估迁移和侵袭能力。通过双荧光素酶报告基因检测和 RNA 免疫沉淀实验证实 miR-29 家族与 PVT1/WAVE1 的相互作用。通过 Western blot 检测 WAVE1、Bcl-2、Bax、cleaved Caspase 3、cyclin D1 和 p21 的蛋白水平。通过异种移植移植实验在体内确定 AML 细胞的致瘤性。AML 患者样本和细胞中 PVT1 表达明显增加，与 WAVE1 表达呈正相关。沉默 PVT1 可抑制 AML 细胞的生长、迁移和侵袭，同时诱导细胞凋亡。此外，PVT1 作为 miR-29 家族的海绵体，增加 AML 细胞中的 WAVE1 表达。在体外和体内异种移植肿瘤中过表达 WAVE1 部分抵消了 PVT1 敲低对 AML 细胞的抗肿瘤作用。PVT1 通过调节 miR-29 家族/WAVE1 轴促进 AML 的进展，这支持了 PVT1 可能是 AML 有前途的治疗靶点的结论。
