上调miRNA-145表达对非小细胞肺癌细胞增殖、凋亡及放射敏感性的影响  

Effect of up regulation of miRNA-145 on cell proliferation, apoptosis and radiosensitivity in non-small cell lung cancer

作　　者：王亚飞[1] 宋长亮[1] 张振军[1] 杨琼[1] 杨庚武[1] 张磊 田云霄[3] 宋小天 WANG Yafei;SONG Changliang;ZHANG Zhenjun;YANG Qiong;YANG Gengwu;ZHANG Lei;TIAN Yunxiao;SONG Xiaotian(Department of Oncology,Handan Central Hospital,Handan 056000,Hebei,China;Department of Orthopedics,Orthopedics Hospital of Handan City,Handan 056000,Hebei,China;Department of Pathology,Handan Central Hospital,Handan 056000,Hebei,China;Department of Immunology,Basic Medical Sciences of Hebei Medical University,Shijiazhuang 050017,Hebei,China)

机构地区：[1]邯郸市中心医院肿瘤科,河北邯郸056000 [2]邯郸市骨科医院骨科,河北邯郸056000 [3]邯郸市中心医院病理科,河北邯郸056000 [4]河北医科大学基础医学院免疫学教研室,石家庄0500170

出　　处：《癌症进展》2019年第10期1152-1155,共4页Oncology Progress

摘　　要：目的探讨上调miRNA-145表达对非小细胞肺癌细胞增殖、凋亡及放射敏感性的影响。方法收集人正常肺上皮细胞株BEAS-2B和非小细胞肺癌细胞株A549,采用逆转录聚合酶链反应(RT-PCR)检测两种细胞中miRNA-145的相对表达量。以Lipofectamine^TM 2000转染试剂将miRNA-145模拟物和阴性对照质粒转染至A549细胞中,分别作为miRNA-145组和NC组,以只加入转染试剂的A549细胞作为对照组,RT-PCR检测各组细胞中miRNA-145的相对表达量。采用四甲基偶氮唑蓝(MTT)法和流式细胞术(FCM)检测上调miRNA-145表达对细胞增殖和凋亡的影响。给予不同放射剂量X线照射后采用克隆形成实验检测细胞的放射敏感性。结果A549细胞中miRNA-145的相对表达量明显低于正常肺上皮BEAS-2B细胞(P<0.01);转染后,miRNA-145组细胞中miRNA-145的相对表达量高于对照组(P<0.05);转染后,NC组与对照组细胞中miRNA-145的相对表达量比较,差异无统计学意义(P﹥0.05);miRNA-145组细胞转染24、48、72 h的光密度(OD)值均低于对照组(P<0.05);转染48 h后,miRNA-145组细胞的凋亡率高于对照组(P<0.05);2、4、6、8 Gy剂量的X线照射后miRNA-145组细胞的存活分数(SF)均低于对照组,放射增敏比(SER)为1.491。结论与正常肺上皮BEAS-2B细胞比较,A549细胞中miRNA-145的相对表达量较低,上调其表达能够抑制A549细胞增殖,促进细胞凋亡,增强放射敏感性。Objective To investigate the effect of up regulation of miRNA-145 on cell proliferation,apoptosis and radiosensitivity in non-small cell lung cancer.Method The human lung epithelial cell line BEAS-2B cells and non-small cell lung cancer A549 cells were collected,and the relative expression level of miRNA-145 was detected by quantitative reverse transcription-polymerase chain reaction(RT-PCR).A549 cells were transfected with the miRNA-145 mimic(miRNA-145 group)and negative control plasmid(NC group)using Lipofectamine^TM2000 transfection reagent,only transfection reagents were added to the control group.The relative expression level of miRNA-145 in above three groups were detected by RT-PCR.Proliferation and apoptosis of A549 cells were also evaluated by methylthiazolyldiphenyl-tetrazolium bromide(MTT)assay and flow cytometry(FCM).The radiosensitivity of the cells was examined by clonogenic assay after various doses of irradiation exposure.Result A549 cells showed significant decreased in miRNA-145 relative expression compared with the normal lung epithelial BEAS-2B cell(P<0.01).After transfection,the relative expression of miRNA-145 in the miRNA-145 group was significant higher than that in the control group(P<0.05),while there was no significant difference between the NC group and the control group(P>0.05).The optical density(OD)values of miRNA-145 group at 24,48,72 h post transfection were lower than those of control group(P<0.05),and the apoptotic rate of nonsmall cell lung cancer cells at 48 h post transfection was higher than that of control group(P<0.05).The cells were irradiated with 2,4,6 or 8 Gy X-ray irradiation and the survival fraction(SF)of cells in the miRNA-145 group was lower than that in the control group,and the enhancement ratio(SER)was 1.491.Conclusion Compared with normal lung epithelial BEAS-2B cells,the relative expression of miRNA-145 is lower in non-small cell lung cancer cells,and up regulation of miRNA-145 can inhibit the proliferation,improve the apoptosis,and enhance the radiosensitivi

关 键 词：非小细胞肺癌 miRNA-145 细胞增殖 细胞凋亡 放射敏感性 

分 类 号：R734.2[医药卫生—肿瘤]