Journal of Prevention and Treatment for Stomatological Diseases ›› 2022, Vol. 30 ›› Issue (6): 381-389.DOI: 10.12016/j.issn.2096-1456.2022.06.001

• Basic Study • Previous Articles     Next Articles

Long non-coding RNA DUXAP9 promotes the proliferation and metastasis of head and neck squamous cell carcinoma

ZHOU Wenkai1,2,3,4,5(), WANG Jiaxuan2, WANG Yuanfeng2, CHEN Meng6, TAO Xingru2, LIU Zheqi1,2,3,4,5, ZHANG Xu1,2,3,4,5, JI Tong1,2,3,4,5(), CAO Wei1,2,3,4,5()   

  1. 1. Department of Oral and Maxillofacial & Head and Neck Oncology, Shanghai Ninth People’s Hospital, Shanghai Jiao Tong University School of Medicine, Shanghai 200011, China
    2. College of Stomatology, Shanghai Jiao Tong University, Shanghai 200011, China
    3. National Center for Stomatology, Shanghai 200011, China
    4. National Clinical Research Center for Oral Diseases, Shanghai 200011, China
    5. Shanghai Key Laboratory of Stomatology, Shanghai 200011, China
    6. College of Public Health, Shanghai Jiao Tong University School, Shanghai 200025, China
  • Received:2021-11-15 Revised:2022-03-04 Online:2022-06-20 Published:2022-04-15
  • Contact: JI Tong, CAO Wei
  • Supported by:
    grants from National Natural Science Foundation of China(81972589)

长链非编码RNA DUXAP9促进头颈鳞癌细胞增殖和转移

周文凯1,2,3,4,5(), 王佳璇2, 王媛凤2, 陈萌6, 陶星如2, 刘喆麒1,2,3,4,5, 张煦1,2,3,4,5, 季彤1,2,3,4,5(), 曹巍1,2,3,4,5()   

  1. 1.上海交通大学医学院附属第九人民医院口腔颌面头颈肿瘤科,上海(200011)
    2.上海交通大学口腔医学院,上海(200011)
    3.国家口腔医学中心,上海(200011)
    4.国家口腔疾病临床医学研究中心,上海(200011)
    5.上海市口腔医学重点实验室,上海(200011)
    6.上海交通大学公共卫生学院,上海(200025
  • 通讯作者: 季彤,曹巍
  • 作者简介:周文凯,医师,硕士,Email: cal310989@sjtu.edu.cn
  • 基金资助:
    国家自然科学基金项目(81972589)

Abstract:

Objective To investigate the role of long non-coding RNA double homeobox A pseudogene 9 (DUXAP9) in head and neck squamous cell carcinoma (HNSCC) and to evaluate the expression level, molecular function and mechanism of DUXAP9 in HNSCC cells. Methods Differential expression of lncRNAs between normal and tumor tissues in HNSCC tissues were screened using lncRNA microarray, the expression level of DUXAP9 in HNSCC tissues and its relationship with prognosis were analyzed in the TCGA database. The expression levels of DUXAP9 in HNSCC tissues and cell lines were detected using qRT-PCR. The function in HNSCC cells after DUXAP9 silencing was evaluated using the CCK-8 assay, wound healing assay, Transwell migration assay and subcutaneous xenograft assay in nude mice. Changes in the transcription and translation of epithelial-mesenchymal transition (EMT)-related proteins in head and neck squamous cell carcinoma cells after DUXAP9 silencing were detected using qRT-PCR and Western blot. Results lncRNA microarray results showed that, compared to adjacent normal tissues, DUXAP9 was abnormally upregulated in HNSCC tissues. Analysis from TCGA database showed that, compared to HNSCC patients with low DUXAP9 expression, HNSCC patients with high DUXAP9 expression had poorer survival. The relative expression of DUXAP9 in HNSCC tissues and 4 HNSCC cell lines increased compared to paired adjacent normal tissues as detected using qRT-PCR. Silencing DUXAP9 significantly inhibited the proliferation, migration and expression of EMT-related genes in HNSCC cells. The silencing of DUXAP9 significantly inhibited subcutaneous tumorigenesis of the HNSCC cell line CAL27 in nude mice. Conclusion Silencing DUXAP9 significantly inhibited the proliferation of HNSCC cells and subcutaneous xenografts in nude mice. DUXAP9 may mediate the migration of head and neck squamous cell carcinoma cells via the EMT pathway.

Key words: long non-coding RNA, double homeobox A pseudogene 9, gene silence, head and neck squamous cell carcinoma, epithelial mesenchymal transformation, E-cadherin, N-cadherin, Vimentin, cell proliferation, cell migration, subcutaneous xenograft in nude mice

摘要:

目的 探讨长链非编码RNA 双同源盒假基因9(double homeobox A pseudogene 9,DUXAP9)在头颈鳞癌中的作用,研究DUXAP9在头颈鳞癌细胞中的表达水平、分子功能和机制。方法 lnc RNA芯片筛选头颈鳞癌组织与癌旁组织差异表达的lncRNA,TCGA数据库中分析DUXAP9在头颈鳞癌组织中的表达水平及其与患者预后的关系;qRT-PCR检测DUXAP9在头颈鳞癌组织样本和细胞系中的表达水平。沉默DUXAP9后,CCK-8实验、细胞划痕实验、Transwell迁移实验和裸鼠皮下成瘤实验评估其在头颈鳞癌细胞中的功能。qRT-PCR和Western blot检测沉默DUXAP9后,头颈鳞癌细胞中上皮-间充质转化(epithelial-mesenchymal transition,EMT)相关蛋白转录及翻译水平的变化。结果 lncRNA芯片结果显示,与癌旁组织相比,DUXAP9在头颈鳞癌中异常高表达。TCGA数据库分析表明,与DUXAP9低表达患者相比,DUXAP9高表达的患者生存率差;qRT-PCR实验表明DUXAP9在头颈鳞癌组织标本和细胞系中异常高表达。沉默DUXAP9可以显著抑制头颈鳞癌细胞的增殖能力、迁移能力和EMT相关基因的表达水平。沉默DUXAP9能显著抑制头颈鳞癌细胞系CAL27的裸鼠皮下成瘤能力。结论 DUXAP9促进头颈鳞癌细胞的增殖、迁移和裸鼠皮下成瘤能力。DUXAP9可能通过调控EMT介导头颈鳞癌细胞的迁移能力。

关键词: 长链非编码RNA, DUXAP9, 基因沉默, 头颈鳞癌, 上皮间质转化, E-钙黏附蛋白, N-钙黏附蛋白, 波形蛋白, 细胞增殖, 细胞迁移, 裸鼠皮下成瘤

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