miR-557通过Notch2/hes1信号通路对肺癌细胞的影响  

Influences of miR-557 on lung cancer cells through Notch2/hesl signaling pathway

作　　者：李国平 周凌燕[1] 刘志聪[1] 於建平 陈凯 张国妹 LI Guoping;ZHOU Lingyan;LIU Zhicong;YU Jianping;CHEN Kai;ZHANG Guomei(Huzhou Central Hospital,Huzhou 313000,China)

机构地区：[1]湖州市中心医院,浙江湖州313000

出　　处：《浙江实用医学》2022年第4期271-277,共7页Zhejiang Practical Medicine

基　　金：浙江省医药卫生科技计划项目(2019RC281)。

摘　　要：目的探讨miR-557通过Notch2/发状分裂相关增强子1(hes1)信号通路对肺癌细胞增殖、侵袭、迁移的影响。方法实时荧光定量PCR(qRT-PCR)法检测人正常肺上皮细胞BEAS-2B及肺癌细胞SPC-A-1、A549、NCI-H446、PC-9中miR-557表达水平。将A549细胞分为对照组(A549细胞在RPMI-1640培养基中正常培养)、miRNC组[A549细胞转染miR-557模拟物(mimic)阴性对照]、miR-557组(A549细胞转染miR-557mimic)、Notch通路激动剂组(5mg/L Jagged1蛋白处理A549细胞)、miR-557+Notch通路激动剂组(转染miR-557mimic成功后加入5mg/L Jagged1蛋白处理A549细胞);qRT-PCR检测A549细胞中miR-557表达水平;细胞计数试剂盒-8检测A549细胞增殖;Transwell检测A549细胞侵袭;划痕实验检测A549细胞迁移;蛋白免疫印迹法检测A549细胞中增殖细胞核抗原(PCNA)、基质金属蛋白酶(MMP)-2、MMP-9、Notch2、hes1蛋白表达水平。结果与人正常肺上皮细胞BEAS-2B比较,肺癌细胞SPC-A-1、A549、NCI-H446、PC-9中miR-557表达水平显著降低(P<0.05),且A549细胞中miR-557表达水平最低,因此,选择A549细胞为研究对象;与对照组、miR-NC组比较,miR-557组A549细胞中miR-557表达水平升高,细胞存活率、侵袭细胞数、划痕愈合率、PCNA、MMP-2、MMP-9、Notch2、hes1蛋白表达水平降低(P<0.05);与对照组比较,Notch通路激动剂组A549细胞中miR-557表达水平差异无统计学意义(P>0.05),细胞存活率、侵袭细胞数、划痕愈合率、PCNA、MMP-2、MMP-9、Notch2、hes1蛋白表达水平升高(P<0.05);与miR-557组比较,miR-557+Notch通路激动剂组A549细胞中miR-557表达水平差异无统计学意义(P>0.05),细胞存活率、侵袭细胞数、划痕愈合率、PCNA、MMP-2、MMP-9、Notch2、hes1蛋白表达水平升高(P<0.05)。结论过表达miR-557通过抑制Notch2/hes1通路抑制A549细胞增殖、侵袭、迁移。Objective To investigate the influences of microRNA-557(miR-557)on the proliferation,invasion and migration of lung cancer cells through the Notch2/hairy and enhancer of split 1(hesl)signaling pathway.Methods Real time fluorescence quantitative PCR(qRT-PCR)was performed to determine the expression levels of miR-557 in human normal lung epithelial cells BEAS-2B and lung cancer cells SPC-A-1,A549,NCI-H446 and PC-9.A549 cells were grouped into control group(A549 cells were normally cultured in RPMI-1640 medium),miR-NC group[A549 cells were transfected with miR-557 mimic negative control],miR-557 group(A549 cells were transfected with miR-557 mimic),Notch pathway agonist group(A549 cells were treated with 5 mg/L Jagged 1 protein),and miR-557+Notch pathway agonist group(after successful transfection of miR-557 mimic,A549 cells were treated with 5 mg/L Jagged 1 protein).qRT-PCR was performed to measure miR-557 expression levels in A549 cells;cell counting kit-8 assay was performed to measure A549 cells proliferation;Transwell was implemented to measure A549 cells invasion;scratch assay was per-fbnned to measure A549 cells migration;Western blot was perfonned to measure proliferating cell nuclear antigen(PCNA),matrix metalloproteinase(MMP)-2,MMP-9,Notch2,hes 1 protein expression levels in A549 cells.Results Compared with human nonnal lung epithelial cells BEAS-2B,the expression levels of miR-557 in SPC-A-1,A549,NCI-H446 and PC-9 decreased significantly(P<0.05),and miR-557 had the lowest expression levels in A549 cells,therefore,A549 cells were selected as the research object;compared with control group and miR-NC group,the expression levels of miR-557 in A549 cells of miR-557 group increased,cell survival rate,number of invasive cells,scratch healing rate,PCNA,MMP-2,MMP-9,Notch2,hesl protein expression levels decreased(P<0.05);compared with control group,there was no significant difference in the expression level of miR-557 in A549 cells of Notch pathway agonist group(P>0.05),the cell survival rate Tnumber of invasive c

关 键 词：miR-557 Notch2/发状分裂相关增强子1通路 肺癌细胞 

分 类 号：R734.2[医药卫生—肿瘤]