Regulation of STAT5 phosphorylation and interaction with SHP1 by lnc-AC004893, a long non-coding RNA overexpressed in myeloproliferative neoplasms.

IF 2 4区医学 Q3 HEMATOLOGY

Hematology Pub Date : 2024-12-01 Epub Date: 2024-07-16 DOI:10.1080/16078454.2024.2375045

Junjun Yang, Jichen Ruan, Bin Zhou, Sisi Ye, Shenmeng Gao, Xiaoqun Zheng

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引用次数: 0

Abstract

Objectives: Constitutive activation of Janus kinase 2 (JAK2)/signal transducer and activator of transcription (STAT) signaling pathway is central to the pathogenesis of myeloproliferative neoplasms (MPNs). Long noncoding RNAs (lncRNAs) regulate diverse biological processes. However, the role of lncRNAs in MPN pathogenesis is not well studied.

Methods: The expression of lnc-AC004893 in MPN patients was measured by quantitative real-time PCR (qRT-PCR). Gene-specific short hairpin RNAs (shRNAs) were designed to inhibit the expression of lnc-AC004893, and western blot was performed to explore the role of lnc-AC004893 via regulating the JAK2/STAT5 signaling pathway. Furthermore, co-IP was performed to determine the binding ability of lnc-AC004893 and STAT5 protein. Finally, the BaF3-JAK2V617F-transplanted mouse model was used to assess the biological role of lnc-ac004893 in vivo.

Results: We report that lnc-AC004893, a poorly conserved pseudogene-209, is substantially upregulated in MPN cells compared with normal controls (NCs). Knockdown of lnc-AC004893 by specific shRNAs suppressed cell proliferation and decreased colony formation. Furthermore, the knockdown of lnc-AC004893 reduced the expression of p-STAT5 but not total STAT5 in HEL and murine IL-3-dependent Ba/F3 cells, which present constitutive and inducible activation of JAK2/STAT5 signaling. In addition, inhibition of murine lnc-ac004893 attenuated BaF3-JAK2V617F-transplanted phenotypes and extended the overall survival. Mechanistically, knockdown of lnc-AC004893 enhanced the binding ability of STAT5 and protein tyrosine phosphatase SHP1. Furthermore, knockdown of lnc-AC004893 decreased STAT5-lnc-AC004893 interaction but not SHP1-lnc-AC004893 interaction.

Conclusion: Lnc-AC004893 regulates STAT5 phosphorylation by affecting the interaction of STAT5 and SHP1. Lnc-AC004893 might be a potential therapeutic target for MPN patients.

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骨髓增殖性肿瘤中过表达的长非编码 RNA lnc-AC004893 对 STAT5 磷酸化及与 SHP1 相互作用的调控

目的：Janus 激酶 2（JAK2）/信号转导和转录激活因子（STAT）信号通路的持续激活是骨髓增殖性肿瘤（MPN）发病机制的核心。长非编码 RNA（lncRNA）调控着多种生物过程。然而，lncRNA在MPN发病机制中的作用还没有得到很好的研究：方法：通过实时定量 PCR（qRT-PCR）检测 MPN 患者体内 lnc-AC004893 的表达。设计了基因特异性短发夹RNA（shRNA）来抑制lnc-AC004893的表达，并进行了Western印迹以探讨lnc-AC004893通过调节JAK2/STAT5信号通路所起的作用。此外，还进行了co-IP检测，以确定lnc-AC004893与STAT5蛋白的结合能力。最后，利用BaF3-JAK2V617F移植小鼠模型评估了lnc-ac004893在体内的生物学作用：结果：我们发现，与正常对照组（NCs）相比，lnc-AC004893（一种保守性很差的假基因-209）在MPN细胞中的表达量大幅上调。通过特异性 shRNA 敲除 lnc-AC004893 可抑制细胞增殖并减少集落形成。此外，敲除 lnc-AC004893 还能降低 HEL 和小鼠 IL-3 依赖性 Ba/F3 细胞中 p-STAT5 的表达，但不能降低总 STAT5 的表达。此外，抑制小鼠lnc-ac004893可减轻BaF3-JAK2V617F移植表型并延长总存活期。从机制上讲，敲除lnc-AC004893增强了STAT5与蛋白酪氨酸磷酸酶SHP1的结合能力。此外，敲除lnc-AC004893会降低STAT5-lnc-AC004893的相互作用，但不会降低SHP1-lnc-AC004893的相互作用：结论：Lnc-AC004893通过影响STAT5和SHP1的相互作用来调节STAT5的磷酸化。Lnc-AC004893可能是MPN患者的潜在治疗靶点。

本文章由计算机程序翻译，如有差异，请以英文原文为准。

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来源期刊

Hematology 医学-血液学

CiteScore

2.60

自引率

5.30%

发文量

140

审稿时长

3 months

期刊介绍： Hematology is an international journal publishing original and review articles in the field of general hematology, including oncology, pathology, biology, clinical research and epidemiology. Of the fixed sections, annotations are accepted on any general or scientific field: technical annotations covering current laboratory practice in general hematology, blood transfusion and clinical trials, and current clinical practice reviews the consensus driven areas of care and management.