miR-582-5p对顺铂耐药鼻咽癌细胞增殖、侵袭与迁移的影响及其机制  

Effect and mechanism of miR-582-5p on proliferation,invasion and migration of cisplatin-resistant nasopharyngeal carcinoma cells

作　　者：刘耘帆 马俊结 苏凯 王晓敏[1] 李慧[1] 马士崟[1] Liu Yunfan;Ma Junjie;Su Kai;Wang Xiaomin;Li Hui;Ma Shiyin(Department of Otolaryngology,the First Affiliated Hospital of Bengbu Medical College,Bengbu 233004,China)

机构地区：[1]蚌埠医学院第一附属医院耳鼻咽喉头颈科,蚌埠233004

出　　处：《中华解剖与临床杂志》2023年第6期400-405,共6页Chinese Journal of Anatomy and Clinics

基　　金：安徽省重点研究与开发计划(202004j07020007);安徽省教育厅科学研究项目(YJS20210540)。

摘　　要：目的探讨miR-582-5p对顺铂耐药鼻咽癌细胞增殖、侵袭与迁移的影响及其机制。方法培养鼻咽癌顺铂耐药细胞株HNE1/DDP,分为对照组和miR-582-5p组;对照组转染miR-582-5p阴性对照试剂,miR-582-5p组转染miR-582-5p模拟物。2组细胞转染后培养48 h,收集细胞采用实时荧光定量聚合酶链反应(qRT-PCR)检测miR-582-5p、苏氨酸激酶3(AKT3)mRNA的表达,细胞计数(CCK)-8实验检测细胞活力,集落形成实验检测细胞增殖能力,划痕实验检测细胞运动能力,Transwell小室检测细胞侵袭及迁移能力,Western blot检测AKT3蛋白的表达情况。结果(1)qRT-PCR检测对照组、miR-582-5p组miR-582-5p mRNA相对表达量分别为1.02±0.08、3.01±0.05,差异有统计学意义(t=38.76,P<0.001)。(2)细胞活力检测结果显示,与对照组比较,miR-582-5p组细胞培养0 h光密度(OD)值差异无统计学意义(P=1.000),培养24、48、72 h时的OD值均降低,差异均有统计学意义(P值均<0.001)。(3)细胞增殖能力检测结果显示,对照组和miR-582-5p组HNE1/DDP细胞菌落数分别是(362.3±13.1)、(155.7±5.0)个,差异有统计学意义(t=25.59,P<0.001)。(4)细胞运动、侵袭、迁移能力检测结果显示,miR-582-5p组细胞迁移率、细胞侵袭数量、细胞迁移数量分别为62.0%±5.0%、(804.3±3.8)个、(631.0±1.0)个,对照组分别为29.3%±4.0%、(434.0±6.1)个、(373.3±7.6)个,差异均有统计学意义(t=8.80、89.53、58.43,P值均<0.001)。(5)miR-582-5p组AKT3 mRNA、蛋白的相对表达量分别为0.99±0.07、1.34±0.02,对照组分别为0.59±0.04、0.60±0.03,差异均有统计学意义(t=8.64、32.03,P值均<0.001)。结论上调miR-582-5p的表达可以抑制顺铂耐药鼻咽癌细胞的增殖、侵袭与迁移能力,其机制可能与下调AKT3的表达有关。Objective To investigate the effect of miR-582-5p on the proliferation,invasion,and migration of cisplatin-resistant nasopharyngeal carcinoma cells and its mechanism.Methods Nasopharyngeal carcinoma HNE1/DDP cells were cultured and divided into control group and miR-582-5p group.The control group was transfected with miR-582-5p negative control reagent,whereas the miR-582-5p group was transfected with miR-582-5p mimics.MiR-582-5p and serine-threonine kinase 3(AKT3)mRNA expression was detected by quantitative real-time polymerase chain reaction(qRT-PCR),cell viability was detected by cell counting kit(CCK-8)assay,cell proliferation was detected by colony formation assay,and cell motility was detected by wound healing assay.The invasion and migration abilities of the cells were detected by Transwell chamber method,AKT3 protein expression was detected by Western blot,and the targeting relationship between miR-582-5p and AKT3 was detected by luciferase reporter experiment.Results(1)The relative mRNA expression levels of miR-582-5p in the control and miR-582-5p groups detected by qRT-PCR were 1.02±0.08 and 3.01±0.05,respectively.Compared with the control group,the relative mRNA expression level of miR-582-5P in the miR-582-5P group increased,and the difference was statistically significant(t=38.76,P<0.001).(2)The results of cell viability test showed that compared with the control group,the optical density(OD)of cell culture in the miR-582-5p group at 0 h was not statistically different(P=1.000).The OD values at 24,48,and 72h decreased,and the differences were statistically significant(all P values<0.001).(3)The results of cell proliferation test showed that the colony number of HNE1/DDP cells in the control and miR-582-5p groups were 362.3±13.1 and 155.7±5.0,respectively;the colony number of HNE1/DDP cells in the miR-582-5p group was significantly lower than that in the control group(t=25.59,P<0.001).(4)The detection results of cell movement,invasion,and migration showed that the cell migration rate,cell invasion

关 键 词：鼻咽肿瘤 抗药性 肿瘤 微小RNA 顺铂 苏氨酸激酶3 细胞增殖 细胞迁移 细胞侵袭 

分 类 号：R739.63[医药卫生—肿瘤]