miR-495对食管癌细胞株Eca109在不同放射剂量和顺铂浓度作用的影响及机制研究  

Effect of miR-495 on Esophageal Cancer Cell Line Eca109 at Different Radiation Doses and Cisplatin Concentrations and Its Mechanism

作　　者：刘伟 周杨 边超 东丽 LIU Wei;ZHOU Yang;BIAN Chao;DONG li(Department of Radiotherapy,Inner Mongolia People’s Hospital,Hohhot 010010,China;Department of Oncology,Inner Mongolia People’s Hospital,Hohhot 010010,China)

机构地区：[1]内蒙古自治区人民医院放射治疗科,呼和浩特010010 [2]内蒙古自治区人民医院肿瘤内科,呼和浩特010010

出　　处：《现代检验医学杂志》2022年第4期13-17,29,共6页Journal of Modern Laboratory Medicine

基　　金：内蒙古自治区自然科学基金(2019MS08090)。

摘　　要：目的探讨微小核糖核酸(microRNA,miR)-495对食管癌细胞株Eca109在不同放射剂量和顺铂浓度作用下的影响及机制。方法采用分次放疗递增法诱导建立放射抵抗型细胞株Eca109(Eca109-RAD)及分次顺铂递增法诱导建立顺铂耐药型细胞株Eca109(Eca109-DDP),实时荧光定量聚合酶链反应(qRT-PCR)检测miR-495在Eca109-RAD及Eca109-DDP细胞中的表达。Eca109-RAD及Eca109-DDP分为NC组和miR-495 mimic组,转染后qRT-PCR检测各组细胞中miR-495的表达,CCK8检测不同放射剂量对Eca109-RAD细胞和Eca109细胞存活能力的影响,及不同浓度的顺铂对Eca109-DDP细胞和Eca109细胞存活能力的影响,NC组和miR-495 mimic组经放射及顺铂处理后,采用流式细胞仪检测各组细胞的凋亡率,Western blot检测各组细胞中凋亡相关蛋白caspase 3,Bax和Bcl-2蛋白的表达。结果与Eca109细胞(0.99±0.01)相比,Eca109-RAD细胞中miR-495的表达(0.48±0.03)及Eca109-DDP细胞中miR-495的表达(0.52±0.05)均降低,差异有统计学意义(t=27.930,15.970,均P=0.000)。与NC组细胞相比,miR-495 mimic组Eca109-RAD细胞中miR-495的表达(4.82±0.48 vs 1.00±0.03)及Eca109-DDP细胞中miR-495的表达(5.68±0.54 vs 1.00±0.01)均显著增加,差异有统计学意义(t=13.760,15.100,均P=0.000)。与NC组相比,miR-495 mimic组Eca109-RAD细胞对放疗敏感度增加,差异均有统计学意义(t=6.780~18.860,P=0.000~0.001);与NC组相比,miR-495 mimic组Eca109-DDP细胞对顺铂敏感度增加,差异均有统计学意义(t=7.510~21.630,均P=0.000)。经放射处理后,与NC组相比,miR-495 mimic组Eca109-RAD细胞平板克隆形成能力(46.33±5.69个vs 93.33±4.51个)及Eca109-DDP(34.67±2.52个vs 89.00±4.03个)细胞平板克隆形成能力显著降低(t=11.210,19.800,均P=0.000)。经放射处理后,与NC组相比,miR-495 mimic组Eca109-RAD细胞凋亡率(25.66%±2.41%vs 8.39%±0.82%)及Eca109-DDP细胞凋亡率(21.05%±5.37%vs 8.67%±1.15%)均显著增加(t=11.750,10.030,P=0.000,0.001)。经放射处理后,与Objective To investigate the effect and mechanism of microRNA(miR)-495 on esophageal cancer cell line Eca109 under different radiation doses and cisplatin concentrations.Methods The radiation resistant cell line Eca109(Eca109 RAD)was induced by fractionated radiotherapy and the cisplatin resistant cell line Eca109(Eca109 DDP)was induced by fractionated cisplatin,the expression of miR-495 in Eca109-RAD and Eca109-DDP cells was detected by real-time fluorescence quantitative polymerase chain reaction(qRT-PCR).Eca109-RAD and Eca109-DDP were divided into NC group and miR-495 mimic group.After transfection,the expression of miR-495 was detected by qRT-PCR,CCK8 was used to detect the effects of different radiation doses on the viability of Eca109 RAD cells and Eca109 cells,and the effects of different concentrations of cisplatin on the viability of Eca109 DDP cells and Eca109 cells.After radiation and cisplatin treatment,the apoptosis rate of cell in NC group and miR-495 mimic group was detected by flow cytometry,and the expressions of apoptosis related proteins caspase 3,Bax and Bcl-2 in each group were detected by Western blot.Results Compared with Eca109 cells(0.99±0.01),the expression of miR-495 in Eca109-RAD cells(0.48±0.03)and Eca109-DDP cells(0.52±0.05)decreased,the differences were statistically significant(t=27.930,15.970,all P=0.000).Compared with NC group,the expression of miR-495 in Eca109-RAD cells(1.00±0.03 vs 4.82±0.48)and Eca109-DDP cells(1.00±0.01 vs 5.68±0.54)in miR-495 mimic group were significantly increased,the differences were statistically significant(t=13.760,15.100,all P=0.000).Compared with NC group,Eca109-RAD cells in miR-495 mimic group were more sensitive to radiotherapy,the differences were statistically significant(t=6.780~18.860,P=0.000~0.0010),and compared with NC group,Eca109-DDP cells in miR-495 mimic group were more sensitive to cisplatin,the differences were statistically significant(t=7.510~21.630,all P=0.000).After radiation treatment,compared with NC group,the plate clone

关 键 词：食管癌 微小核糖核酸-495 凋亡 放化疗敏感度 

分 类 号：R735.1[医药卫生—肿瘤] R730.43[医药卫生—临床医学]