摘要
背景与目的:miR-1290可以调节干扰素调节因子-2(interferon regulatory factor-2,IRF2)的表达,调节非小细胞肺癌(non-small cell lung cancer,NSCLC)的恶性生物学行为,但其具体作用机制目前尚不清楚,因此,探讨miR-1290靶向IRF2对NSCLC细胞增殖、侵袭的调控作用及相关机制。方法:将体外培养的A549细胞分为miR-1290 mimic组、miR-1290 inhibitor组和NC组;实时荧光定量聚合酶链反应(real-time fluorescence quantitative polymerase chain reaction,RTFQ-PCR)和蛋白质印迹法(Western blot)检测各组细胞miR-1290和IRF2的表达,采用荧光素酶实验进一步验证靶向关系。将体外培养的A549细胞分为NC组(转染空白质粒)、miR-1290组(转染miR-1290)、IRF2组(转染IRF2)和miR-1290+IRF2组(转染miR-1290+IRF2);细胞计数试剂盒(cell counting kit-8,CCK-8)检测各组细胞的增殖活性;Transwell实验检测各组细胞的侵袭,划痕实验检测细胞迁移,Western blot检测各组细胞血管内皮生长因子(vascular endothelial growth factor,VEGF)、E-钙黏着蛋白(E-cadherin)、基质金属蛋白酶(matrix metalloproteinase,MMP)-2和Ki-67。结果:miR-1290可以靶向负调控IRF2的表达;与NC组相比,miR-1290组miR-1290的表达明显上调(P<0.05),IRF2的表达明显下调(P<0.05),细胞增殖活性、侵袭力和迁移率明显上调(P<0.05),VEGF、MMP-2和Ki-67明显上调(P<0.05),E-cadherin水平明显下调(P<0.05),IRF2组IRF2的表达明显上调(P<0.05),细胞增殖活性、侵袭力和迁移率明显下调(P<0.05),VEGF、MMP-2和Ki-67明显下调(P<0.05),E-cadherin水平明显上调(P<0.05);与IRF2组相比,miR-1290+IRF2组miR-1290的表达明显上调(P<0.05),IRF2的表达明显下调(P<0.05),细胞增殖活性、侵袭力和迁移率明显上调(P<0.05),VEGF、MMP-2和Ki-67明显上调(P<0.05),E-cadherin水平明显下调(P<0.05)。结论:miR-1290可能通过靶向负调控IRF2的表达,促进NSCLC细胞的增殖和侵袭。
Background and purpose:miR-1290 can regulate the expression of interferon regulatory factor-2(IRF2)and the malignant biological behavior of non-small cell lung cancer(NSCLC),but the specific mechanism of its action is still unclear.Therefore,this study explored the regulatory effect of miR-1290 targeting IRF2 on the proliferation and invasion of NSCLC cells and its related mechanism.Methods:A549 cells cultured in vitro were divided into miR-1290 mimic group,miR-1290 inhibitor group and NC group.Real-time fluorescence quantitative polymerase chain reaction(RTFQ-PCR)and Western blot were performed to detect expression of miR-1290 and IRF2 in each group.Their targeted relationship was further verified by fluorescence assay.A549 cells cultured in vitro were divided into NC group(transfected with mimic),miR-1290 group(transfected with miR-1290),IRF2 group(transfected with IRF2)and miR-1290+IRF2 group(transfected with miR-1290+IRF2).Cell counting kit-8(CCK-8)was performed to detect cell proliferation activity in each group.Transwell assay was performed to detect cell invasion in each group.The cell migration was detected by scratch test.The levels of vascular endothelial growth factor(VEGF),E-cadherin,matrix metalloproteinase(MMP)-2 and Ki-67 in each group were detected by Western blot.Results:IRF2 expression could be negatively regulated by miR-1290.Compared with NC group,miR-1290 expression was significantly up-regulated in miR-1290 group(P<0.05),while IRF2 expression was significantly down-regulated(P<0.05).Moreover,cell proliferation activity,invasion ability and migration rate were significantly increased(P<0.05),levels of VEGF,MMP-2 and Ki-67 were significantly up-regulated(P<0.05),and expression of E-cadherin was significantly down-regulated(P<0.05)in miR-1290 group.IRF2 expression was significantly upregulated in IRF2 group(P<0.05),cell proliferation activity,invasion ability and migration rate were significantly decreased(P<0.05),levels of VEGF,MMP-2 and Ki-67 were significantly down-regulated(P<0.05),and protein level of E-cadherin was significantly up-regulated(P<0.05).Compared with IRF2 group,miR-1290 expression was significantly up-regulated in miR-1290+IRF2 group,while expression of IRF2 was significantly down-regulated(P<0.05).In addition,cell proliferation activity,invasion ability and migration rate were significantly increased(P<0.05),levels of VEGF,MMP-2 and Ki-67 were significantly up-regulated(P<0.05),and protein level of E-cadherin was significantly down-regulated(P<0.05).Conclusion:MiR-1290 may promote proliferation and invasion of NSCLC cells by negatively regulating IRF2 expression.
作者
董磊
张仲妍
DONG Lei;ZHANG Zhongyan(Department of Oncology,Wuhan Red Cross Hospital,Wuhan 430000,Hubei Province,China;Department of Oncology and Traditional Chinese Medicine,Oncology Department,Chongqing University Affiliated Tumor Hospital,Chongqing 400030,China)
出处
《中国癌症杂志》
CAS
CSCD
北大核心
2020年第1期32-40,共9页
China Oncology
