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《天津医科大学学报》[ISSN:1006-8147/CN:12-1259/R]

卷:

32卷

期数:

2026年01期

页码:

6-13,22

栏目:

肿瘤疾病专题

出版日期:

2026-01-20

文章信息/Info

Title:

LncRNA HOTAIRM1 promotes the progression of acute myeloid leukemia by interacting with the ENO1 protein

文章编号:

1006-8147(2026)01-0006-09

作者:

刘秋岑1; 张祝琴2; 饶书权1

（1.天津医科大学细胞生态海河实验室，天津 300020；2.中国医学科学院基础医学研究所，北京协和医学院基础学院，医学分子生物学国家重点实验室生物化学与分子生物学系，北京 100005）

Author(s):

LIU Qiucen1;  ZHANG Zhuqin2;  RAO Shuquan1

（1.Haihe Laboratory of Cell Ecosystem， Tianjin Medical University， Tianjin 300020，China； 2.Department of Molecular Biology and Biochemistry， State Key Laboratory of Medical Molecular Biology， Institute of Basic Medical Sciences CAMS， Chinese Academy of Medical Sciences， School of Basic Medicine， Peking Union Medical College， Beijing 100005， China）

关键词:

急性髓系白血病; 长链非编码RNA; HOX反义基因间RNA髓系1; α-烯醇化酶; NEDD4L

Keywords:

acute myeloid leukemia;  long non-coding RNA;  HOX antisense intergenic RNA myeloid 1;  α-Enolase;  NEDD4L

分类号:

R737.31

DOI:

10.20135/j.issn.1006-8147.2026.01.0006

文献标志码:

A

摘要:

目的：探讨链长非编码RNA HOX反义基因间RNA髓系1（HOTAIRM1）在急性髓系白血病（AML）中的生物学功能及其分子机制。方法：利用生物信息学分析方法筛选出在AML中高表达的lncRNA；GEPIA2数据库验证HOTAIRM1在AML中的表达情况；Kaplan-Meier生存分析评估HOTAIRM1在AML中的临床预后价值。将HL-60、THP-1细胞分为si-NC组和si-HOTAIRM1组，通过qRT-PCR验证小干扰RNA干扰效率，结合CCK-8和流式细胞实验观察HOTAIRM1对AML细胞增殖和凋亡表型的影响。根据RNA pull-down和质谱鉴定后生信分析结果筛选HOTAIRM1潜在的互作蛋白；HDOCK分子对接验证HOTAIRM1与目标蛋白α-烯醇化酶（ENO1）的结合。Western印迹检测HOTAIRM1对ENO1表达的影响；小干扰RNA联合CHX抑制剂处理AML细胞后通过Western印迹观察HOTAIRM1对ENO1蛋白稳定性的调控作用；利用UbiBrowser数据库预测可能修饰ENO1泛素化的E3泛素连接酶；免疫共沉淀（Co-IP）实验检测在AML细胞中NEDD4L与ENO1的相互结合，观察HOTAIRM1与ENO1、NEDD4L之间的相互作用。结果：生信分析发现，AML患者高表达HOTAIRM1；在GEPIA2数据库中验证HOTAIRM1呈高表达（P<0.05），且HOTAIRM1的高表达与临床预后不良密切相关（P<0.05）。与对照组相比，用针对HOTAIRM1的小干扰RNA处理HL-60和THP-1细胞有显著的敲低效率（t=9.28、10.23，均P<0.05）；敲低HOTAIRM1可抑制HL-60、THP-1细胞增殖（t=6.53、4.78，均P<0.05），并促进HL-60和THP-1细胞凋亡（t=18.54、15.63，均P<0.001）。生信分析RNA pull-down后的质谱结果，筛选出与HOTAIRM1潜在的互作蛋白ENO1；HDOCK分子对接验证了HOTAIRM1与蛋白ENO1具有较强的结合活性。敲低HOTAIRM1后能够下调HL-60和THP-1细胞ENO1的蛋白表达水平（t=15.37、17.92，均P<0.001），且在CHX抑制剂作用下以时间依赖的方式逐渐降解（t=12.95、10.23，均P<0.01）。NEDD4L作为E3泛素连接酶介导ENO1蛋白的泛素化修饰，且ENO1、NEDD4L两种蛋白能在AML细胞内相互结合。敲低HOTAIRM1的AML细胞中NEDD4L与ENO1之间的相互作用增强，HL-60和THP-1细胞总体泛素化水平升高（t=4.72、6.58，均P<0.05）。结论：LncRNA HOTAIRM1在AML中呈高表达，且通过抑制NEDD4L介导的ENO1泛素化降解从而促进AML进展。

Abstract:

Objective： To investigate the biological function and molecular mechanism of long non-coding RNA HOX antisense intergenic RNA myeloid 1 （lncRNA HOTAIRM1） in acute myeloid leukemia （AML）. Methods： Using bioinformatics analysis methods， the lncRNAs that are highly expressed in AML were screened out. GEPIA2 database was used to verify the expression of HOTAIRM1 in AML， and Kaplan-Meier survival analysis was used to evaluate the clinical prognostic value of HOTAIRM1 in AML. The HL-60 and THP-1 cells were divided into the si-NC group and the si-HOTAIRM1 group， and the interference efficiency of the small interfering RNA was verified by qRT-PCR. CCK-8 and flow cytometry were used to observe the effect of HOTAIRM1 on the proliferation and apoptotic phenotype of AML cells. Potential interacting proteins of HOTAIRM1 were screened based on bioinformatics analysis of RNA pull-down and mass spectrometry identification results. The binding of HOTAIRM1 to the target protein α-Enolase （ENO1） was verified by HDOCK molecular docking. The effect of HOTAIRM1 on ENO1 expression was detected by Western blotting. The regulation of HOTAIRM1 on ENO1 protein stability was observed by Western blotting after AML cells were treated with small interfering RNA combined with CHX inhibitors. E3 ubiquitin ligases that might modify ENO1 ubiquitination were predicted using the UbiBrowser database. The mutual binding of NEDD4L and ENO1 in AML cells and the interaction between HOTAIRM1 and ENO1， NEDD4L were observed by co-immunoprecipitation （Co-IP） assay. Results： Based on Bioinformatics analysis， it was found that HOTAIRM1 was highly expressed in AML patients. High expression of HOTAIRM1 was verified in the GEPIA2 database （P<0.05）， and its high expression was closely related to poor clinical prognosis （P<0.05）. Compared with the control group， small interfering RNA targeting HOTAIRM1 showed remarkable knockdown efficiency in HL-60 and THP-1 cells （t=9.28， 10.23， both P<0.05）. Knocking down HOTAIRM1 inhibited HL-60 and THP-1 cells proliferation （t=6.53， 4.78， both P<0.05）， and promoted apoptosis （t=18.54， 15.63， both P<0.001）. Bioinformatics analysis of mass spectrometry after RNA pull-down was used to screen the potential interaction protein ENO1 with HOTAIRM1. HDOCK molecular docking verified that HOTAIRM1 had strong binding activity with the protein ENO1. Knockdown of HOTAIRM1 was able to downregulate ENO1 protein expression levels in HL-60 and THP-1 cells （t=15.37， 17.92， both P<0.001）， and gradually degraded in a time-dependent manner in response to CHX inhibitors （t=12.95， 10.23， both P<0.01）. NEDD4L acted as an E3 ubiquitin ligase to mediate ubiquitination modification of ENO1 protein. Both ENO1 and NEDD4L proteins could bind to each other in AML cells. The interaction between NEDD4L and ENO1 in AML cells was enhanced with knocked-down HOTAIRM1， and the overall ubiquitination level increased in HL-60 and THP-1 cells （t=4.72， 6.58， both P<0.05）. Conclusion： LncRNA HOTAIRM1 is highly expressed in AML and promotes the AML progression by inhibiting NEDD4L-mediated ubiquitylation degradation of ENO1.

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备注/Memo

备注/Memo:

基金项目:京津冀基础研究合作专项（J230018）
作者简介:刘秋岑（2000-），女，硕士在读，研究方向：血液病学；
通信作者:饶书权，E-mail：raoshuquan@ihcams.ac.cn。

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更新日期/Last Update: 2026-01-15