华西口腔医学杂志 ›› 2024, Vol. 42 ›› Issue (1): 46-55.doi: 10.7518/hxkq.2024.2023237

• 基础研究 • 上一篇    下一篇

miR-362-3p调控垂体肿瘤转化基因1抑制口腔鳞状细胞癌侵袭及增殖的研究

丁啸1,2(), 陈嘉雯2, 曲鹏宇1, 孙晨雨1, 李洪利3, 胡温庭1, 范欣1()   

  1. 1.潍坊医学院附属医院口腔科,潍坊 261000
    2.潍坊医学院口腔医学院,潍坊 261053
    3.潍坊医学院医学研究实验中心,潍坊 261053
  • 收稿日期:2023-07-26 修回日期:2023-12-12 出版日期:2024-02-01 发布日期:2024-01-12
  • 通讯作者: 范欣 E-mail:dx6666yy@126.com;fanxinsunny@aliyun.com
  • 作者简介:丁啸,住院医师,硕士,E-mail:dx6666yy@126.com
  • 基金资助:
    山东省自然科学基金项目(ZR202110190030);潍坊医学院附属医院种子基金项目(2021wffyzzjj06);潍坊市科技发展计划项目(2022YX029)

miR-362-3p inhibited the invasion and metastasis of oral squamous cell carcinoma cells by targeting the regulation of pituitary tumor-transforming gene 1

Ding Xiao1,2(), Chen Jiawen2, Qu Pengyu1, Sun Chenyu1, Li Hongli3, Hu Wenting1, Fan Xin1()   

  1. 1.Dept. of Stomatology, Affiliated Hospital of Weifang Medical University, Weifang 261000, China
    2.School of Stomatology, Weifang Medical University, Weifang 261053, China
    3.Medical Research Center, Weifang Medical University, Weifang 261053, China
  • Received:2023-07-26 Revised:2023-12-12 Online:2024-02-01 Published:2024-01-12
  • Contact: Fan Xin E-mail:dx6666yy@126.com;fanxinsunny@aliyun.com
  • Supported by:
    Natural Science Foundation Project of Shandong Province(ZR202110190030);Fundamental Research Program Funding of Affiliated Hospital of Weifang Medical University(2021wffyzzjj06);Weifang Science and Technology Development Plan Project(2022YX029);Correspondence: Fan Xin, E-mail: fanxinsunny@aliyun.com

摘要:

目的 探讨垂体肿瘤转化基因1(PTTG1)在miR-362-3p作用下对口腔鳞状细胞癌(OSCC)细胞Cal-27、HN-30侵袭以及增殖能力的影响。 方法 生物信息学在线数据库查询PTTG1在头颈部鳞状细胞癌(HNSCC)中的表达。蛋白质印迹法(Western blot)实验检测PTTG1在Cal-27、HN-30以及HOK细胞系中的表达。划痕愈合实验、Transwell侵袭实验及5-乙炔基-2’脱氧尿嘧啶核苷(EdU)细胞增殖实验检测PTTG1对Cal-27、HN-30细胞迁移、侵袭、增殖的影响。生物信息学在线数据库预测PTTG1的上游miRNA,双荧光素酶实验检测结合情况,实时荧光定量聚合酶链反应(qRT-PCR)检测该miRNA在组织中的表达。 结果 ENCORI数据库结果显示PTTG1在OSCC组织中表达上调;Western blot实验显示Cal-27、HN-30细胞中PTTG1表达量较HOK细胞中高。转染Si-PTTG1质粒的Cal-27、HN-30细胞的迁移能力、侵袭能力和细胞增殖能力均较对照组降低(P<0.05)。通过网站预测出PTTG1的上游miRNA为miR-362-3p,双荧光素酶实验检测出PTTG1与miR-362-3p存在结合位点;qRT-PCR检测结果显示miR-362-3p在OSCC肿瘤组织中相对于正常组织表达下调(P<0.05);并且敲低miR-362-3p的表达后能够促进敲低PTTG1后的Cal-27、HN-30侵袭和增殖。 结论 miR-362-3p可通过靶向PTTG1抑制Cal-27、HN-30细胞侵袭、增殖。

关键词: 口腔鳞状细胞癌, 垂体肿瘤转化基因1, 微小RNA, 侵袭, 增殖

Abstract:

Objective This study aimed to explore the effect of pituitary tumor-transforming gene 1 (PTT-G1) on the invasion and proliferation of oral squamous cell carcinoma (OSCC) cell lines under the action of miR-362-3p. Methods The bioinformatics online database was used to query the expression of PTTG1 in head and neck squamous cell carcinoma (HNSCC). The expression of PTTG1 in the Cal-27, HN-30, and HOK cell lines was detected by Western blot. A wound-healing assay was used to determine the effect of PTTG1 on the migration ability of the OSCC cells. The Transwell assay was used to examine the changes in cell-invasion ability. 5-ethynyl-2'-deoxyuridine (EdU) cell-proliferation assay was used to detect changes in cell-proliferation ability. Bioinformatics approach predicted the upstream miRNA of PTTG1. The targeting relationship between miR-362-3p and PTTG1 was examined by the dual luciferase assay, and quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the expression of miRNA in OSCC tissues. Results The ENCORI database showed that PTTG1 expression was up-regulated in OSCC tissues. Western blot confirmed that PTTG1 expression was up-regulated in Cal-27 and HN-30 cells than HOK cells. PTTG1 knockout can inhibit the migration, invasion, and proliferation of Cal-27 and HN-30 cells (P<0.05). Bioinformatics prediction websites predicted that the upstream miRNA of PTTG1 was miR-362-3p, and PTTG1 can bind to miR-362-3p. Results of qRT-PCR showed that miR-362-3p expression was downregulated in OSCC tissues compared with normal tissue (P<0.05). Transwell and EdU experiments confirmed that miR-362-3p knockdown can promote the invasion and proliferation of Cal-27 and HN-30 after PTTG1 knockdown. Conclusion miR-362-3p can inhibit the invasion and proliferation of Cal-27 and HN-30 cells by targeting PTTG1.

Key words: oral squamous cell carcinoma, pituitary tumor-transforming gene 1, microRNA, invasion, proliferation

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