摘要
目的探讨miR-590-3p通过调控Dicer1对鼻咽癌细胞侵袭转移的影响及其作用机制。方法收集2017年8月-2019年2月于我院经手术切除且被证实为鼻咽癌的组织标本以及对应的癌旁组织标本各40例,RTFQ-PCR检测miR-590-3p在鼻咽癌组织及癌旁组织、人鼻黏膜上皮细胞(HNEpC)、鼻咽癌细胞株CNE-1、CNE-1-mimics-miR-590-3p及阴性对照CNE-1-mimics-NC中的表达情况。Western blot检测Dicer1在各细胞株中的表达。生物信息学预测及双荧光素酶报告基因实验验证miR-590-3p和Dicer1的靶向关系。Transwell检测miR-590-3p对鼻咽癌细胞侵袭能力的影响;划痕实验检测miR-590-3p对鼻咽癌细胞迁移能力的影响。IFA检测miR-590-3p对上皮间质转化(EMT)相关分子N-cadherin表达的影响。结果miR-590-3p在鼻咽癌组织和鼻咽癌细胞株CNE-1中的表达水平分别低于癌旁组织和正常人鼻黏膜上皮细胞;与CNE-1及CNE-1-mimics-NC相比,稳定过表达细胞系CNE-1-mimics-miR-590-3p中miR-590-3p、Dicer1的表达水平均显著升高(P<0.05)。经Target Scan数据库分析发现miR-590-3p和Dicer1有互补结合位点,双荧光素酶报告基因实验证实两者能靶向结合。过表达miR-590-3p能显著降低鼻咽癌细胞的侵袭及转移能力,同时抑制N-cadherin的表达水平。结论miR-590-3p在鼻咽癌中通过靶向上调Dicer1的表达,广泛促进miRNA的成熟,降低N-cadherin的表达,进而抑制鼻咽癌细胞的EMT进程以及侵袭转移能力。
Objective To investigate the effect of miR-590-3 p on the invasion and metastasis of nasopharyngeal carcinoma cells via regulating Dicer1.Methods From August 2017 to February 2019,40 cases of nasopharyngeal carcinoma tissue and 40 cases of paracancerous tissue samples were surgically removed in our hospital and collected.RTFQ-PCR was used to detect the expression of miR-590-3 p.The expression of Dicer1 in each cell line was detected by Western blot.Bioinformatics prediction and dual-luciferase reporter gene assay verified the targeting relationship between miR-590-3 p and Dicer1.Transwell assay was performed to detect the effect of miR-590-3 p on the invasion ability of nasopharyngeal carcinoma cells.Scratch test was performed to detect the effect of miR-590-3 p on the migration ability of nasopharyngeal carcinoma cells.IFA detected the effect of miR-590-3 p on the expression of Ncadherin,an EMT-related molecule.Results The expression levels of miR-590-3 p were lower in nasopharyngeal carcinoma tissue and cell line CNE-1 than in paracancerous tissue and normal human nasal mucosa epithelial cell HNEpC.Compared with CNE-1 and CNE-1-mimics-NC,the expression levels of miR-590-3 p and Dicer1 in the stably overexpressed cell line CNE-1-mimics-miR-590-3 p were significantly higher(P<0.05).Target Scan database analysis revealed that miR-590-3 p and Dicer1 had complementary binding sites,and dual-luciferase reporter gene experiments confirmed that both can bind to the Target.Overexpression of miR-590-3 p could significantly reduce the invasion and metastasis of nasopharyngeal carcinoma cells and inhibit the expression of N-cadherin.Conclusion miR-590-3 p can extensively promote the maturation of nasopharyngeal carcinoma miRNA and reduce the expression of N-cadherin by targeting the up-regulated expression of Dicer1,inhibiting the EMT process and invasion and metastasis of nasopharyngeal carcinoma cells.
作者
刘建光
王张锋
高国钰
LIU Jian-guang;WANG Zhang-feng;GAO Guo-yu(Department of Otolaryngology,Hebei Provincial Chest Hospital,Shijiazhuang 050048;Department of Otolaryngology,Kailuan General Hospital,Tangshan 063000,China)
出处
《解剖科学进展》
2020年第5期544-549,共6页
Progress of Anatomical Sciences
