DDX6通过调控CKMT1A mRNA稳定性促进鼻咽癌细胞增殖和迁移的机制研究

熊茄妍; 雷伟; 游波; 张振新; 谢海静; 单颖; 夏天; 周勇

doi:10.19401/j.cnki.1007-3639.2024.05.002

您当前的位置：

首页 >

文章列表页 >

DDX6通过调控CKMT1A mRNA稳定性促进鼻咽癌细胞增殖和迁移的机制研究

论著 | 更新时间：2025-12-31

- DDX6通过调控CKMT1A mRNA稳定性促进鼻咽癌细胞增殖和迁移的机制研究
- Study on the mechanism of DDX6 promoting proliferation and migration of nasopharyngeal carcinoma cells by regulating stability of CKMT1A mRNA
- 中国癌症杂志 2024年34卷第5期页码：451-459
- 作者机构：
  
  1. 南通大学附属医院耳鼻咽喉头颈外科研究所，江苏南通 226001
  2. 南通大学医学院，江苏南通 226001
  3. 苏州大学附属第二医院耳鼻咽喉头颈外科，江苏苏州 215000
  4. 南通大学附属医院耳鼻咽喉头颈外科，江苏南通 226001
- 作者简介：
  
  [ "熊茄妍（ORCID: 0009-0007-8213-9178），硕士。" ]
  周勇（ORCID: 0000-0002-5928-8489），硕士，主治医师。
- 基金信息：
  
  国家自然科学基金(82103435);国家自然科学基金(82203799)
- DOI：10.19401/j.cnki.1007-3639.2024.05.002
  中图分类号： R739.6
- 收稿：2023-11-01，
  
  修回：2024-02-21，
  
  纸质出版：2024-05-30
- 稿件说明：
移动端阅览
熊茄妍, 雷伟, 游波, 等. DDX6通过调控CKMT1A mRNA稳定性促进鼻咽癌细胞增殖和迁移的机制研究[J]. 中国癌症杂志, 2024,34(5):451-459.

Jiayan XIONG, Wei LEI, Bo YOU, et al. Study on the mechanism of DDX6 promoting proliferation and migration of nasopharyngeal carcinoma cells by regulating stability of CKMT1A mRNA[J]. China Oncology, 2024, 34(5): 451-459.
熊茄妍, 雷伟, 游波, 等. DDX6通过调控CKMT1A mRNA稳定性促进鼻咽癌细胞增殖和迁移的机制研究[J]. 中国癌症杂志, 2024,34(5):451-459. DOI： 10.19401/j.cnki.1007-3639.2024.05.002.

Jiayan XIONG, Wei LEI, Bo YOU, et al. Study on the mechanism of DDX6 promoting proliferation and migration of nasopharyngeal carcinoma cells by regulating stability of CKMT1A mRNA[J]. China Oncology, 2024, 34(5): 451-459. DOI： 10.19401/j.cnki.1007-3639.2024.05.002.

摘要

背景与目的：

DDX是一类腺苷三磷酸（adenosine triphosphate，ATP）依赖的RNA解旋酶，与mRNA调控、肿瘤增殖及侵袭等密切相关。本研究旨在探讨DDX家族成员DDX6对CKMT1A mRNA稳定性的影响以及DDX6-CKMT1A轴对人鼻咽癌细胞CNE2增殖和迁移能力的影响及其分子机制。

方法：

检索癌症基因组图谱（The Cancer Genome Atlas，TCGA）数据库中DDX6和CKMT1A在人头颈部鳞状细胞癌中的表达情况并进行相关性分析，利用蛋白质印迹法（Western blot）检测南通大学附属医院保存的人体鼻咽癌组织和正常鼻咽部组织中CKMT1A和DDX6的表达，

本研究通过了南通大学附属医院伦理委员会的审查（编号：2022-L114）。利用transwell实验检测细胞迁移能力，采用EdU试剂盒检测细胞增殖能力，采用细胞集落形成实验检测克隆形成能力。转染慢病毒、质粒，构建南通大学附属医院保存的人源鼻咽癌细胞CNE2的sh-DDX6、sh-CKMT1A、sh-CKMT1A+sh-DDX6和oe-CKMT1A细胞模型，明确DDX6和CKMT1A表达水平对鼻咽癌细胞恶性生物学表型的影响。构建BALC/c裸小鼠皮下移植瘤模型，检测DDX6和CKMT1A在小鼠体内对鼻咽癌细胞成瘤性的影响。采用RNA稳定性实验检测敲除DDX6对CKMT1A mRNA的影响，进一步明确DDX6的分子机制。

结果：

人鼻咽癌组织中DDX6高表达，CKMT1A低表达，DDX6与CKMT1A表达呈负相关。DDX6通过破环CKMT1A mRNA稳定性，抑制CKMT1A蛋白质翻译。在CNE2细胞中，CKMT1A低表达可增强细胞迁移和增殖能力，高表达则抑制细胞迁移和增殖能力，而DDX6的敲除可逆转CKMT1A下调导致的恶性行为进展。在裸鼠皮下移植瘤模型中，低表达CKMT1A促进肿瘤细胞生长，低表达DDX6抑制肿瘤细胞生长，而同时敲除DDX6与CKMT1A能恢复单独敲低DDX6导致的抑制效果。

结论：

鼻咽癌细胞中DDX6通过破坏CKMT1A mRNA稳定性，负调控CKMT1A蛋白质翻译，增强鼻咽癌细胞增殖和迁移能力，从而促进鼻咽癌恶性进展。

Abstract

Background and purpose:

DDX is an adenosine triphosphate (ATP)-dependent RNA helicase closely related to mRNA regulation

tumor proliferation and invasion. This article aimed to explore the effect of DDX6

a member of the DDX family

on the stability of CKMT1A mRNA

as well as the effect of the DDX6 CKMT1A axis on the proliferation and migration ability of human nasopharyngeal carcinoma cell CNE2 and its molecular mechanism.

Methods:

We retrieved the data of expressions of DDX6 and CKMT1A in human head and neck squamous cell carcinoma from The Cancer Genome Atlas (TCGA) database and performed a correlation analysis. Western blot was performed to detect the expressions of CKMT1A and DDX6 in human nasopharyngeal carcinoma tissues and normal nasopharyngeal tissues preserved by Affiliated Hospital of Nantong University. This study was approved by the Ethics Committee of Affiliated Hospital of Nantong University (Number: 2022-L114). We used transwell assay to detect cell migration ability

EdU assay to detect cell proliferation ability

and colony formation assay to detect clone formation ability. We transfect with lentivirus and plasmids to construct sh-DDX6

sh-CKMT1A

sh-CKMT1A+sh-DDX6 and oe-CKMT1A cell models derived from the human nasopharyngeal carcinoma ce

ll line CNE2

preserved by Affiliated Hospital of Nantong University

to clarify the impact of DDX6 and CKMT1A expression levels on the malignant biological phenotypes of nasopharyngeal carcinoma cells. BALB/c nude mice subcutaneous xenograft tumor model was constructed to detect the effects of DDX6 and CKMT1A on nasopharyngeal carcinoma cells in mice. RNA stability assay was used to detect the effect of DDX6 knockout on CKMT1A mRNA and further clarify the molecular mechanism of DDX6.

Results:

DDX6 was highly expressed

CKMT1A level was low in human nasopharyngeal carcinoma tissue

and DDX6 was negatively correlated with CKMT1A expression. DDX6 inhibited protein translation of CKMT1A by disrupting its mRNA stability. Low expression of CKMT1A in CNE2 cells enhanced cell migration and proliferation ability

while high expression inhibited migration and proliferation ability. Knocking out DDX6 reversed the progression of malignant behavior caused by downregulation of CKMT1A. Low expression of CKMT1A promoted tumor cell growth in BALB/cnude mice subcutaneous xenograft tumor model

while low expression of DDX6 inhibited tumor cell growth. Knocking out DDX6 and CKMT1A simultaneously restored the inhibitory effect caused by knocking down DDX6 alone.

Conclusion:

DDX6 in nasopharyngeal carcinoma cells disrupts the stability of CKMT1A mRNA

negatively regulates CKMT1A protein translation

upregulates the proliferation and migration ability of nasopharyngeal carcinoma cells

and promotes malignant progression of nasopharyngeal carcinoma.

关键词

Keywords

references

YOKOTA T , HOMMA A , KIYOTA N , et al. Immunotherapy for squamous cell carcinoma of the head and neck [J ] . Jpn J Clin Oncol , 2020 , 50 ( 10 ): 1089 - 1096 . DOI: 10.1093/jjco/hyaa139 http://doi.org/10.1093/jjco/hyaa139

MO Y Z , WANG Y M , WANG Y A , et al. Circular RNA circPVT1 promotes nasopharyngeal carcinoma metastasis via the β-TrCP/c-Myc/SRSF1 positive feedback loop [J ] . Mol Cancer , 2022 , 21 ( 1 ): 192.

LU D , YUNIS J J . Cloning, expression and localization of an RNA helicase gene from a human lymphoid cell line with chromosomal breakpoint 11q23.3 [J ] . Nucleic Acids Res , 1992 , 20 ( 8 ): 1967 - 1972 .

BOURGEOIS C F , MORTREUX F , AUBOEUF D . The multiple functions of RNA helicases as drivers and regulators of gene expression [J ] . Nat Rev Mol Cell Biol , 2016 , 17 ( 7 ): 426 - 438 .

TAJIRIKA T , TOKUMARU Y , TANIGUCHI K , et al. DEAD-box protein RNA-helicase DDX6 regulates the expression of HER2 and FGFR2 at the post-transcriptional step in gastric cancer cells [J ] . Int J Mol Sci , 2018 , 19 ( 7 ): 2005 .

GHASHGHAEI M , LE C T , SHAALAN H , et al. miR-148a-3p and DDX6 functional link promotes survival of myeloid leukemia cells [J ] . Blood Adv , 2023 , 7 ( 15 ): 3846 - 3861 .

FAN H X , LV P , MU T , et al. LncRNA n335586/miR-924/CKMT1A axis contributes to cell migration and invasion in hepatocellular carcinoma cells [J ] . Cancer Lett , 2018 , 429 : 89 - 99 . DOI: S0304-3835(18)30335-5 http://doi.org/S0304-3835(18)30335-5

YANG M J , WANG X C , YE Z H , et al. Mitochondrial creatine kinase 1 regulates the cell cycle in non-small cell lung cancer via activation of cyclin-dependent kinase 4 [J ] . Respir Res , 2023 , 24 ( 1 ): 111.

SHI H D , SONG Y L , SONG Z , et al. CKMT1B is a potential prognostic biomarker and associated with immune infiltration in Lower-grade glioma [J ] . PLoS One , 2021 , 16 ( 1 ): e0245524.

AMAMOTO R , UCHIUMI T , YAGI M , et al. The expression of ubiquitous mitochondrial creatine kinase is downregulated as prostate cancer progression [J ] . J Cancer , 2016 , 7 ( 1 ): 50 - 59 . DOI: 10.7150/jca.13207 http://doi.org/10.7150/jca.13207

李刚 . CKMT1A对人鼻咽癌HONE1细胞铂类化疗药物敏感性的影响及机制研究 [J ] . 中南医学科学杂志 , 2019 , 47 ( 6 ): 576 - 580 .

LI G . Mitochondrial creatine kinase 1A in sensitivity of nasopharyngeal carcinoma cells to platinum-based chemotherapeutics [J ] . Med Sci J Cent South China , 2019 , 47 ( 6 ): 576 - 580 .

COLAPRICO A , SILVA T C , OLSEN C , et al. TCGAbiolinks: an R/Bioconductor package for integrative analysis of TCGA data [J ] . Nucleic Acids Res , 2016 , 44 ( 8 ): e71.

HAAS R C , KORENFELD C , ZHANG Z F , et al. Isolation and characterization of the gene and cDNA encoding human mitochondrial creatine kinase [J ] . J Biol Chem , 1989 , 264 ( 5 ): 2890 - 2897 .

SU C H , TANG Y D , ZHENG C F . DExD/H-box helicases: multifunctional regulators in antiviral innate immunity [J ] . Cell Mol Life Sci , 2021 , 79 ( 1 ): 2.

HEERMA VAN VOSS M R , VAN DIEST P J , RAMAN V . Targeting RNA helicases in cancer: the translation trap [J ] . Biochim Biophys Acta Rev Cancer , 2017 , 1868 ( 2 ): 510 - 520 .

VRIES S D , NAARMANN-DE VRIES I S , URLAUB H , et al. Identification of DEAD-box RNA helicase 6 (DDX6) as a cellular modulator of vascular endothelial growth factor expression under hypoxia [J ] . J Biol Chem , 2013 , 288 ( 8 ): 5815 - 5827 . DOI: 10.1074/jbc.M112.420711 http://doi.org/10.1074/jbc.M112.420711

AKAO Y , MATSUMOTO K , OHGUCHI K , et al. Human DEAD-box/RNA unwindase rck/p54 contributes to maintenance of cell growth by affecting cell cycle in cultured cells [J ] . Int J Oncol , 2006 , 29 ( 1 ): 41 - 48 .

TANIGUCHI K , IWATSUKI A , SUGITO N , et al. Oncogene RNA helicase DDX6 promotes the process of c-Myc expression in gastric cancer cells [J ] . Mol Carcinog , 2018 , 57 ( 5 ): 579 - 589 .

DU Z , YUAN J H , WU Z W , et al. Circulating exosomal circRNA_0063476 impairs expression of markers of bone growth via the miR-518c-3p/DDX6 axis in ISS [J ] . Endocrinology , 2022 , 163 ( 11 ): bqac138.

ZHAO Z Y , WU C J , HE X Y , et al. miR-152-3p aggravates vascular endothelial cell dysfunction by targeting DEAD-box helicase 6 (DDX6) under hypoxia [J ] . Bioengineered , 2021 , 12 ( 1 ): 4899 - 4910 . DOI: 10.1080/21655979.2021.1959864 http://doi.org/10.1080/21655979.2021.1959864

SCHWARZENBACH H , GAHAN P B . Interplay between lncRNAs and microRNAs in breast cancer [J ] . Int J Mol Sci , 2023 , 24 ( 9 ): 8095.

ZHOU C , HE T D , CHEN L J . LncRNA CASC19 accelerates chondrocytes apoptosis and proinflammatory cytokine production to exacerbate osteoarthritis development through regulating the miR-152-3p/DDX6 axis [J ] . J Orthop Surg Res , 2021 , 16 ( 1 ): 399.

浏览量

1928

下载量

CSCD

文章被引用时，请邮件提醒。

提交

工具集

关联资源

SEC14L1P1对口腔鳞状细胞癌细胞增殖和迁移的影响

长链非编码RNA FLJ30679对口腔鳞状细胞癌细胞增殖和迁移的影响

GOLM1调控PI3K/AKT/mTOR信号转导通路促进肺腺癌细胞增殖、侵袭和迁移的机制研究

PTP1B基因对人胃癌细胞株的增殖和迁移能力的影响

RNA干扰YAP基因对人膀胱癌T24细胞株增殖和迁移能力的影响