miR-148a-3p调控EGFR抑制涎腺腺样囊性癌细胞的增殖、侵袭和转移
miR-148a-3p inhibits the proliferation,invasion and metastasis of human salivary adenoid cystic carcino-ma cells by targeting EGFR
高万鹏 1赵琦 2惠琪 2王嘉乐 2郭嘉飞 2杨子桧 2王珺 2魏建华 2杨新杰2
作者信息
- 1. 154000,佳木斯大学附属口腔医学院·附属口腔医院黑龙江省口腔生物医学材料及临床应用重点实验室
- 2. 口颌系统重建与再生全国重点实验室,国家口腔疾病临床医学研究中心,陕西省口腔疾病临床医学研究中心,空军军医大学第三附属医院口腔颌面头颈肿瘤科
- 折叠
摘要
目的:研究miR-148a-3p对涎腺腺样囊性癌SACC-LM细胞的增殖、侵袭和转移的影响及相关分子机制.方法:将miR-148a-3p模拟物和抑制剂,靶向EGFR的siRNA以及相应对照转染SACC-LM细胞.生物信息学手段分析预测miR-148a-3p的潜在靶基因;qRT-PCR检测miR-148a-3p和EGFR的mRNA表达;Western blot实验检测EGFR的蛋白表达.CCK-8、划痕和Transwell分别用于检测SACC-LM细胞的增殖、迁移和侵袭能力;双荧光素酶报告基因检测miR-148a-3p与EGFR之间的直接靶向关系;SPSS 22.0软件对数据进行统计分析.结果:过表达或抑制miR-148a-3p可以显著抑制或促进SACC-LM细胞的增殖、侵袭和转移(P＜O.05);生物信息学分析和双荧光素酶实验显示miR-148a-3p可靶向调控EGFR的表达(P＜0.001);下调EGFR可以显著抑制SACC-LM细胞的增殖、迁移和侵袭能力(P＜0.05),并可部分逆转miR-148a-3p抑制剂对这些能力的促进作用(P＜0.05).结论:涎腺腺样囊性癌细胞中miR-148a-3p的下调导致其靶基因EGFR的异常高表达,从而促进了涎腺腺样囊性癌细胞的增殖、侵袭和转移.
Abstract
Objective:To study the effects and the related molecular mechanisms of miR-148a-3p on the proliferation,invasion and migration of salivary adenoid cystic carcinoma SACC-LM cells.Methods:miR-148a-3p mimics and inhibitors,siRNA targeting EG-FR and their corresponding controls were transfected into SACC-LM cells.Bioinformatics was used to predict the potential target genes of miR-148a-3p.EGFR and miR-148a-3p mRNA expression levels were examined by qRT-PCR and the protein levels of EG-FR were detected by Western blotting.CCK-8,scratch,and Transwell assays were used to study the proliferation,migration,and invasion of SACC-LM cells,respectively.The direct targeting relationship between miR-148a-3p and EGFR was examined by using the double luciferase reporter gene assay.Statistical analysis of the data was performed by SPSS 22.0 software.Results:Overexpres-sion or inhibition of miR-148a-3p significantly inhibited or promoted the proliferation,invasion and metastasis of SACC-LM cells re-spectively(P＜0.05).Bioinformatics and double luciferase assay showed that miR-148a-3p directly targeted and regulated the expres-sion of EGFR(P＜0.001).Downregulation of EGFR inhibited the proliferation,migration and invasion of SACC-LM cells(P＜0.05)and partially reversed the promoting effect of miR-148a-3p inhibition(P＜0.05).Conclusion:The downregulation of miR-148a-3p leads to the abnormally high expression of its target gene EGFR,and promotes the proliferation,invasion,and migration of salivary adenoid cystic carcinoma cells.
关键词
涎腺腺样囊性癌/miR-148a-3p/EGFR/增殖/侵袭/转移Key words
Adenoid cystic carcinoma of sali-vary gland/miR-148a-3p/EG-FR/Proliferation/Invasion/Mi-gration引用本文复制引用
基金项目
国家自然科学基金(82173165)
国家自然科学基金青年基金(82002867)
国家口腔疾病临床医学研究中心专项(LCB202008)
出版年
2024
NETL
NSTL
万方数据