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

doi:10.7518/hxkq.2024.2023237

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

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.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.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/Abstract

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

中图分类号:

丁啸, 陈嘉雯, 曲鹏宇, 孙晨雨, 李洪利, 胡温庭, 范欣. 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[J]. 华西口腔医学杂志, 2024, 42(1): 46-55.

Ding Xiao, Chen Jiawen, Qu Pengyu, Sun Chenyu, Li Hongli, Hu Wenting, Fan Xin. miR-362-3p inhibited the invasion and metastasis of oral squamous cell carcinoma cells by targeting the regulation of pituitary tumor-transforming gene 1[J]. West China Journal of Stomatology, 2024, 42(1): 46-55.

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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

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

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