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 regula-tion of pituitary tumor-transforming gene 1
丁啸 1陈嘉雯 2曲鹏宇 3孙晨雨 3李洪利 4胡温庭 3范欣3
作者信息
- 1. 潍坊医学院附属医院口腔科,潍坊 261000;潍坊医学院口腔医学院,潍坊 261053
- 2. 潍坊医学院口腔医学院,潍坊 261053
- 3. 潍坊医学院附属医院口腔科,潍坊 261000
- 4. 潍坊医学院医学研究实验中心,潍坊 261053
- 折叠
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.
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 prolif-eration 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.
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Key words
oral squamous cell carcinoma/pituitary tumor-transforming gene 1/microRNA/invasion/prolif-eration引用本文复制引用
基金项目
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)
出版年
2024
国家科技期刊平台
NSTL
万方数据