microRNA98靶向N-RAS对鼻咽癌CNE-1细胞增殖与迁移的影响

Effects of microRNA98 targeting N-RAS on the proliferation and migration of nasopharyngeal carcinoma CNE-1 cells

目的探讨miR-98对鼻咽癌CNE-1细胞增殖与迁移的影响及可能机制。方法利用脂质体介导的miR-98模拟物或阴性对照转染鼻咽癌CNE-1细胞,并通过实时定量PCR进行验证。MTT实验检测CNE-1细胞增殖能力的变化,划痕实验检测CNE-1迁移能力变化,采用生物信息学软件预测N-RAS是否为miR-98潜在的靶基因,并以双荧光素酶报告基因实验验证。Western blot检测miR-98过表达后CNE-1细胞N-RAS蛋白的表达变化以及下游ERK、p-ERK、AKT、p-AKT的表达变化。结果 MiR-98过表达能抑制鼻咽癌CNE-1细胞的体外增殖与迁移能力;生物信息学软件分析结果显示N-RAS是miR-98的靶基因之一,双荧光素酶报告基因证实N-RAS为miR-98的下游靶基因。miR-98过表达后,CNE-1细胞中N-RAS表达下调,ERK与Akt蛋白的表达水平不变,但pERK与p-Akt蛋白的表达水平下调。结论 miR-98通过靶向调控N-RAS抑制鼻咽癌CNE-1细胞增殖与迁移。

Objective To investigate the effect and possible mechanism of microRNA-98 on the proliferation and migration of nasopharyngeal carcinoma CNE-1 cells.Methods NPC CNE-1 cells were transfected with liposomemediated mimics of microRNA-98 or negative control,and verified by real-time quantitative PCR.MTT assay was used to detect the proliferation of CNE-1 cells,scratch assay was used to detect the migration of CNE-1 cells,and bioinformatics software was used to predict whether N-RAS was a potential target gene of microRNA-98,which was evaluated by dual luciferase reporter assay.Western blot was used to detect the expression of N-RAS protein and downstream ERK,p-ERK,AKT and p-AKT proteins in CNE-1 cells after over-expression of microRNA-98.Results MiR-98 overexpression inhibited the proliferation and migration of nasopharyngeal carcinoma CNE-1 cells in vitro.Bioinformatics software analysis showed that N-RAS was one of the target genes of microRNA-98,and double luciferase reporter assay confirmed that NRAS was the downstream target gene of microRNA-98.After the overexpression of microRNA-98,the expression level of NRAS was down-regulated in CNE-1 cells,while the expression levels of ERK and Akt proteins remained unchanged,but the expression levels of p-ERK and p-Akt protein were down-regulated.Conclusion Mi-98 inhibited the proliferation and migration of nasopharyngeal carcinoma CNE-1 cells by targeting N-RAS.