长链非编码 RNA HOXC-AS3 通过稳定 KDM5B 加速宫颈癌癌细胞的恶性增殖。

LncRNA HOXC-AS3 accelerates malignant proliferation of cervical cancer cells via stabilizing KDM5B.

机构信息

Department of Obstetrics and Gynecology, Suzhou Wuzhong People's Hospital, Jiangsu Province, No. 61, Dongwu North Road, Suzhou City, 215128, China.

出版信息

J Cancer Res Clin Oncol. 2024 Jun 6;150(6):294. doi: 10.1007/s00432-024-05799-y.

DOI:10.1007/s00432-024-05799-y

PMID:38842683

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC11156713/

Abstract

BACKGROUND

Cervical cancer (CC) is a common malignancy amongst women globally. Ubiquitination plays a dual role in the occurrence and development of cancers. This study analyzed the mechanism of long noncoding RNA HOXC cluster antisense RNA 3 (lncRNA HOXC-AS3) in malignant proliferation of CC cells via mediating ubiquitination of lysine demethylase 5B (KDM5B/JARID1B).

METHODS

The expression patterns of lncRNA HOXC-AS3 and KDM5B were measured by real-time quantitative polymerase chain reaction or Western blot analysis. After transfection with lncRNA HOXC-AS3 siRNA and pcDNA3.1-KDM5B, proliferation of CC cells was assessed by the cell counting kit-8, colony formation, and 5-Ethynyl-2'-deoxyuridine staining assays. The xenograft tumor model was established to confirm the impact of lncRNA HOXC-AS3 on CC cell proliferation in vivo by measuring tumor size and weight and the immunohistochemistry assay. The subcellular location of lncRNA HOXC-AS3 and the binding of lncRNA HOXC-AS3 to KDM5B were analyzed. After treatment of lncRNA HOXC-AS3 siRNA or MG132, the protein and ubiquitination levels of KDM5B were determined. Thereafter, the interaction and the subcellular co-location of tripartite motif-containing 37 (TRIM37) and KDM5B were analyzed by the co-immunoprecipitation and immunofluorescence assays.

RESULTS

LncRNA HOXC-AS3 and KDM5B were upregulated in CC tissues and cells. Depletion of lncRNA HOXC-AS3 repressed CC cell proliferation and in vivo tumor growth. Mechanically, lncRNA HOXC-AS3 located in the nucleus directly bound to KDM5B, inhibited TRIM37-mediated ubiquitination of KDM5B, and upregulated the protein levels of KDM5B. KDM5B overexpression attenuated the inhibitory role of silencing lncRNA HOXC-AS3 in CC cell proliferation in vivo and in vitro.

CONCLUSION

Nucleus-located lncRNA HOXC-AS3 facilitated malignant proliferation of CC cells via stabilization of KDM5B protein levels.

摘要

背景

宫颈癌（CC）是全球女性中常见的恶性肿瘤。泛素化在癌症的发生和发展中起着双重作用。本研究通过介导赖氨酸去甲基酶 5B（KDM5B/JARID1B）的泛素化来分析长非编码 RNA HOXC 簇反义 RNA 3（lncRNA HOXC-AS3）在 CC 细胞恶性增殖中的机制。

方法

实时定量聚合酶链反应或 Western blot 分析检测 lncRNA HOXC-AS3 和 KDM5B 的表达模式。转染 lncRNA HOXC-AS3 siRNA 和 pcDNA3.1-KDM5B 后，通过细胞计数试剂盒-8、集落形成和 5-乙炔基-2'-脱氧尿苷染色测定 CC 细胞的增殖。通过测量肿瘤大小和重量以及免疫组织化学测定，建立异种移植肿瘤模型，以体内验证 lncRNA HOXC-AS3 对 CC 细胞增殖的影响。分析 lncRNA HOXC-AS3 的亚细胞定位和 lncRNA HOXC-AS3 与 KDM5B 的结合。用 lncRNA HOXC-AS3 siRNA 或 MG132 处理后，测定 KDM5B 的蛋白和泛素化水平。此后，通过 co-immunoprecipitation 和免疫荧光测定分析三肽重复含 37 （TRIM37）和 KDM5B 的相互作用和亚细胞共定位。

结果

lncRNA HOXC-AS3 和 KDM5B 在 CC 组织和细胞中上调。沉默 lncRNA HOXC-AS3 抑制 CC 细胞增殖和体内肿瘤生长。机制上，lncRNA HOXC-AS3 位于细胞核内，直接与 KDM5B 结合，抑制 TRIM37 介导的 KDM5B 泛素化，上调 KDM5B 蛋白水平。KDM5B 过表达减弱了沉默 lncRNA HOXC-AS3 对 CC 细胞体内和体外增殖的抑制作用。