miR-145-5p通过调节PHB2基因发挥对非小细胞肺癌细胞A549增殖迁移的影响  

miR-145-5p Influences the Proliferation and Migration of Non-Small Cell Lung Cancer Cells by Regulating PHB2

作　　者：李新军 钟敏华[1] 吴展陵[1] 曹玲莉[1] 邱琦 涂平华 汪娜娜 彭闪闪 高飞 LI Xinjun;ZHONG Minhua;WU Zhanling(Xiaogan Central Hospital,Hubei Xiaogan 432000,China)

机构地区：[1]湖北省孝感市中心医院,湖北孝感432000

出　　处：《河北医学》2021年第9期1418-1423,共6页Hebei Medicine

基　　金：湖北省科技计划项目,(编号:2017FFB6320)。

摘　　要：目的:探讨miR-145-5p通过靶向调控PHB2基因抑制非小细胞肺癌细胞(NSCLC)的增殖和迁移的作用及机制。方法:使用RT-PCR法评估miR-145-5p在NSCLC组织和癌旁组织正常人支气管上皮细胞系(BEAS-2B)和非小细胞肺癌细胞系(A549)中的表达;MTT及EDU测定miR-145-5p过表达后A549细胞的活力;划痕实验和Transwell评估miR-145-5p过表达对NSCLC迁移和侵袭的影响;荧光素酶报告基因测定法确定miR-145-5p靶向调控PHB2蛋白编码基因PHB2 mRNA的3'-非翻译区(3'-UTR)发挥作用;蛋白质免疫印迹法检测A549细胞中PHB2蛋白表达;pcDNA-PHB2转染A549细胞,并评估细胞增殖、迁移和侵袭的水平。结果:miR-145-5p在NSCLC与癌旁组织相比呈现明显低表达(P<0.01),约为癌旁组织的47.54±10.12%;利用miR-145-5p mimic转染细胞后,可在体外显著抑制肿瘤细胞增殖、迁移和侵袭(P<0.001),分别约为对照组的28.31±13.97%、30.57±5.84%、32.56±6.99%;miR-145-5p通过直接结合PHB2 mRNA的3'-UTR发挥抑制作用,从而负面调节PHB2的表达;miR-145-5p和PHB2共转染A549细胞后,其增殖、迁移和侵袭能力较只过表达miR-145-5p组明显增强。结论:miR-145-5p通过靶定PHB2 mRNA来抑制非小细胞肺癌细胞的增殖、迁移和侵袭,并且可能是抗癌治疗中有前景的治疗方向。Objective:To explore the role and mechanism of miR-145-5p in regulating the proliferation and migration of non-small cell lung cancer cells by targeting PHB2.Methods:The expression of miR-145-5p in NSCLC tissues and adjacent tissues,normal human bronchial epithelial cell line(BEAS-2B),and non-small cell lung cancer cell line(A549)was assessed by RT-PCR.The effect of miR-145-5p overexpression on NSCLC activity was determined by MTT and EDU.The effect of miR-145-5p overexpression on migration and invasion of NSCLC was evaluated by scratch test and Transwell.The luciferase reporter assay confirmed the role of miR-145-5p in the 3'-untranslated region(3'-UTR)of the PHB2 protein coding gene PHB2 mRNA.The expression of PHB2 protein in A549 cells was detected by western blot.A549 cells were transfected with pcDNA-phB2 and the levels of cell proliferation,migration and invasion were assessed.Results:The expression of miR-145-5p in NSCLC was significantly lower than that in paracancerous tissues(P<0.01),which was about 47.54±10.12%of paracancerous tissues.After transfection with miR-145-5p mimic,the proliferation,migration and invasion of tumor cells were significantly inhibited in vitro(P<0.001),which were about 28.31±13.97%,30.57±5.84%and 32.56±6.99%of those in the control group,respectively.miR-145-5p negatively regulates the expression of PHB2 by directly binding the 3'-UTR of PHB2 mRNA.After co-transfection with miR-145-5p and PHB2,the proliferation,migration and invasion ability of A549 cells were significantly enhanced compared with that of miR-145-5p group alone.Conclusion:MiR-506-3p inhibitions the proliferation,migration and invasion of non-small cell lung cancer cells by targeting PHB2 mRNA,and may be a promising therapeutic direction in anti-cancer therapy.

关 键 词：miR-145-5p PHB2 非小细胞肺癌 

分 类 号：R734.2[医药卫生—肿瘤]