摘要
目的:探讨微小RNA-429(miR-429)在鼻咽癌细胞中的表达及其对细胞增殖、迁移及侵袭能力的影响。方法:选取本院收治的78例鼻咽癌患者为鼻咽癌组,另选取同期因其他疾病需进行鼻内镜手术患者33例为对照组。采用实时荧光定量PCR(qRT-PCR)法检测两组组织中miR-429的表达。体外培养人鼻咽上皮细胞NP69与鼻咽癌细胞株CNE-2,将miR-429阴性对照质粒、miR-429 mimics质粒分别转染至CNE-2细胞,分别命名为阴性转染组、miR-429过表达组,未经处理的细胞命名为空白组,采用qRT-PCR技术检测各转染组细胞中miR-429表达。MTT法检测各转染组细胞增殖情况;平板克隆细胞形成实验检测细胞菌落形成情况;Transwell检测细胞迁移及侵袭能力;蛋白免疫印迹(Western blot)检测E盒结合锌指蛋白(ZEB1)、基质金属蛋白酶2(MMP-2)、钙黏附蛋白E(E-cadherin)蛋白表达量。结果:鼻咽癌组织中miR-429表达水平显著低于对照组(P<0.05),且CNE-2细胞中miR-429表达水平显著低于NP69细胞(P<0.05);miR-429过表达组miR-429表达水平显著高于空白组、阴性转染组(P<0.05);miR-429过表达组细胞增殖能力显著低于空白组、阴性转染组(P<0.05);miR-429过表达组细胞菌落形成、迁移细胞及侵袭细胞数量均显著低于空白组、阴性转染组(P<0.05);miR-429过表达组细胞中ZEB1、MMP-2蛋白表达水平显著低于空白组、阴性转染组,而E-cadherin蛋白表达水平显著升高(P <0.05)。结论:鼻咽癌细胞中miR-429呈低表达,miR-429过表达后可显著抑制鼻咽癌细胞增殖、迁移及侵袭。
Objective:To investigate the expression of microRNA-429(miR-429)in nasopharyngeal carcinoma cells and its effects on cell proliferation,migration and invasion.Methods:78 patients with nasopharyngeal carcinoma were selected as nasopharyngeal carcinoma group,and 33 patients who underwent endoscopic sinus surgery for other diseases were selected as control group.The expression of miR-429 in tissues of two groups was detected by real-time fluorescence quantitative PCR(qRT-PCR).Human nasopharyngeal epithelial cell line NP69 and nasopharyngeal carcinoma cell line CNE-2 were cultured in vitro.miR-429 negative control plasmid and miR-429 mimics plasmid were transfected into CNE-2 cells and named as negative transfection group and miR-429 overexpression group respectively.The untreated cells were named blank group.The expression of miR-429 in cells of each transfected group was detected by qRT-PCR.MTT assay was used to detect the proliferation of each transfection group.Cell colony formation was detected by plate clone cell formation experiment.Transwell was used to detect cell migration and invasion.The expressions of E-box binding zinc finger protein(ZEB1),matrix metalloproteinase-2(MMP-2)and calcium adhesion protein E(E-cadherin)were detected by Western blot.Results:The expression of miR-429 in nasopharyngeal carcinoma tissues was significantly lower than that in control group(P<0.05),and the expression of miR-429 in CNE-2 cells was significantly lower than that in NP69 cells(P<0.05).The expression level of miR-429 in miR-429 overexpression group was significantly higher than that in blank group and negative transfection group(P<0.05).The proliferation ability of miR-429 overexpression group was significantly lower than that of blank group and negative transfection group(P<0.05).The numbers of colonies,migrating cells and invasive cells in the overexpression group were significantly lower than those in the blank group and negative transfection group(P<0.05).The expressions of ZEB1 and MMP-2 proteins in the overexpression group were significantly lower than those in the blank group and negative transfection group,while the expression of E-cadherin protein was significantly higher(P<0.05).Conclusion:The expression of miR-429 in nasopharyngeal carcinoma is low,overexpression of miR-429 can significantly inhibit the proliferation,migration and invasion of nasopharyngeal carcinoma cells.
作者
谭建成
刘鑫国
邢金燕
TAN Jiancheng;LIU Xinguo;XING Jinyan(Department of Otolaryngology,Zhumadian Central Hospital,Henan Zhumadian 463000,China)
出处
《现代肿瘤医学》
CAS
北大核心
2021年第9期1501-1506,共6页
Journal of Modern Oncology
