肺癌miR-146a表达及其对细胞增殖、侵袭及迁移的影响  

Expression of miR-146a in lung cancer and its effect on cell proliferation,invasion and migration

作　　者：陈辉国 周亚夫[1] 颜建华[1] 肖国华 田双如 杨劲松[1] CHEN Huiguo;ZHOU Yafu;YAN Jianhua;XIAO Guohua;TIAN Shuangru;YANG Jinsong(Department of Cardio-Thoracic Surgery,Hunan Provincial People s Hospital,the First Affiliated Hospital of Hunan Normal University,Changsha 410005,China)

机构地区：[1]湖南省人民医院湖南省师范大学附属第一医院心胸外科,长沙410005

出　　处：《临床肿瘤学杂志》2020年第3期224-229,共6页Chinese Clinical Oncology

摘　　要：目的探讨微小RNA-146a(miR-146a)在肺癌组织和细胞中的表达和甲基化状态,及其对A549细胞增殖、侵袭、迁移的影响及可能机制。方法收集2018年1月至2019年4月在我院行根治性手术的非小细胞肺癌组织和对应癌旁组织,实时荧光定量PCR(QPCR)和甲基化特异性PCR(MSP)检测miR-146a的表达水平和甲基化状态,并分析甲基化状态与肺癌临床病理特征的关系。采用5-氮杂-2’-脱氧胞苷(5-AZA-2’-dC)处理A549细胞(处理组),MTT、Transwell实验和划痕实验检测处理组细胞增殖、侵袭和迁移活性。采用Western blotting检测Notch1和发状分裂相关增强子1(Hes-1)蛋白表达。结果肺癌组织和A549细胞中miR-146a表达量分别为0.63±0.28、0.85±0.11,均低于癌旁正常组织和BEAS-2B细胞(P<0.05)。MSP检测显示肺癌组织miR-146a甲基化率为62.5%(50/80),高于癌旁组织(P<0.05);miR-146a甲基化状态与肿瘤直径、TNM分期、淋巴结转移有关(P<0.05)。处理组细胞miR-146a表达水平为2.15±0.48,高于空白对照组(P<0.05);处理组细胞增殖、侵袭和迁移活性均低于空白对照组(P<0.05)。处理组Notch1蛋白和Hes-1蛋白表达水平分别为0.24±0.05和0.22±0.06,均低于空白对照组(P<0.05)。结论启动子异常甲基化导致在肺癌组织和细胞中miR-146a表达量降低,可能通过减弱对Notch1/Hes-1信号通路的抑制作用,促进肺癌细胞增殖、侵袭和转移,miR-146a有望成为肺癌新的生物治疗靶点。Objective To investigate the expression and methylation of microRNA-146a(miR-146a)in lung cancer tissues and cells,and its effect on the proliferation,invasion and migration of A549 cells.Methods The non-small cell lung cancer tissues and the corresponding paracarcinoma tissues were collected from January 2018 to April 2019 for radical operation in our hospital.The expression level and methylation status of miR-146a were detected by real-time fluorescence quantitative PCR(QPCR)and methylation specific PCR(MSP).The relationship between methylation status and clinicopathological characteristics of patients was analyzed.A549 cells(treatment group)were treated with 5-Aza-2’-deoxycytidine(5-Aza-2’-dC).The proliferation,invasion and migration activities of A549 cells in treatment group were detected by MTT,Transwell and scratch tests.Western blotting was used to detect the protein expression of Notch 1 and Hes-1.Results The expression levels of miR-146a in lung cancer and A549 cells were 0.63±0.28 and 0.85±0.11,respectively,which were lower than those in normal tissues and BEAS-2B cells(P<0.05).The methylation rate of miR-146a in lung cancer was 62.5%(50/80),which was higher than that in adjacent tissues(P<0.05).The methylation of miR-146a was related to tumor diameter,TNM stage and lymph node metastasis(P<0.05).The expression level of miR-146a in the treatment group was 2.15±0.48,which was higher than that in the blank control group(P<0.05);the proliferation,invasion and migration activities in the treatment group were lower than that in the blank control group(P<0.05).The expression levels of Notch1 and Hes-1 in the treatment group were 0.24±0.05 and 0.22±0.06,respectively,lower than that in the control group(P<0.05).Conclusion The abnormal methylation of promoter results in the decrease of miR-146a expression in lung cancer tissues and cells,which may promote the proliferation,invasion and metastasis of lung cancer cells by reducing the inhibition of Notch 1/Hes-1 signaling pathway,and miR-146a may become a

关 键 词：非小细胞肺癌 MIR-146A Notch1通路 甲基化 

分 类 号：R734.2[医药卫生—肿瘤]