基于NF-κB/MAPK信号通路miR-125a-5p诱导乳腺癌MCF7细胞自噬的作用机制研究  

Mechanism of miR-125a-5p inducing autophagy in breast cancer MCF7 cells based on NF-κB/MAPK signaling pathway

作　　者：郭建美[1] 杨颖博[2] 李杰[3] GUO Jianmei;YANG Yingbo;LI Jie(Department of Oncology,Baoding Municipal First Central Hospital,Baoding,Hebei 071000,China;First Department of General Medicine,Baoding Municipal First Central Hospital,Baoding,Hebei 071000,China;Department of Breast Surgery,Fourth Hospital of Hebei Medical University,Shijiazhuang,Hebei 050000,China)

机构地区：[1]河北省保定市第一中心医院肿瘤内科,071000 [2]河北省保定市第一中心医院全科医疗一科,071000 [3]河北医科大学第四医院乳腺外科,石家庄050000

出　　处：《重庆医学》2022年第1期20-24,28,共6页Chongqing medicine

基　　金：河北省医学科学研究计划项目(20201105)。

摘　　要：目的探讨微小RNA-125a-5p(miR-125a-5p)基于核因子-κB(NF-κB)/有丝分裂原活化蛋白激酶(MAPK)信号通路诱导乳腺癌MCF7细胞自噬的作用机制。方法取对数期生长乳腺癌MCF7细胞,分为miR-125a-5p组、miR-NC组和空白组,采用慢病毒转染法将hsa-miR-125a-5p mimics、NC-mimics分别转染至miR-125a-5p组和miR-NC组,空白组不给予处理。采用实时荧光定量聚合酶链反应(RT-qPCR)检测转染后各组细胞miR-125a-5p基因相对表达水平;采用单丹磺酰二胺及Hoechst双染色检测各组细胞自噬情况,并采用Western blot法检测自噬相关蛋白Beclin-1、微管相关蛋白1轻链3-Ⅱ(LC3Ⅱ)表达情况;采用RT-qPCR、Western blot法检测各组细胞NF-κB/MAPK信号通路相关基因及蛋白相对表达水平。结果空白组、miR-NC组、miR-125a-5p组miR-125a-5p基因相对表达水平分别为0.44±0.08、0.42±0.07、1.27±0.16,miR-125a-5p组miR-125a-5p基因相对表达水平高于空白组和miR-NC组,差异有统计学意义(P<0.05);空白组、miR-NC组、miR-125a-5p组细胞阳性率分别为(2.10±0.52)%、(2.30±0.47)%、(26.40±7.14)%,miR-125a-5p组细胞阳性率高于空白组和miR-NC组,差异有统计学意义(P<0.05);miR-125a-5p组自噬相关蛋白Beclin1、LC3Ⅱ蛋白相对表达水平高于空白组和miR-NC组,差异有统计学意义(P<0.05)。3组NF-κB p65、细胞外调节蛋白激酶(ERK1/2)、MAPK p38 mRNA相对表达水平比较,差异均无统计学意义(P>0.05);miR-125a-5p组NF-κB/MAPK信号通路磷酸化蛋白与相应蛋白比值p-NF-κB p65/NF-κB p65、p-ERK1/2/ERK1/2、p-MAPK p38/MAPK p38均高于空白组和miR-NC组,差异均有统计学意义(P<0.05)。结论过表达miR-125a-5p基因可诱导乳腺癌MCF7细胞发生自噬,可能与NF-κB/MAPK信号通路激活有关。Objective To investigate the mechanism of microRNA(miR)-125a-5p-induced autophagy in breast cancer MCF7 cells based on nuclear factor(NF)-κB and mitogen-activated protein kinase(MAPK)signaling pathways.Methods MCF7 cells with logarithmic growth were divided into the miR-125a-5p group,miR-NC group and blank group.Hsa-miR-125a-5p mimics and NC mimics were transfected into miR-125a-5p group and miR-NC group by lentivirus transfection.The blank group was not treated.The relative expression of miR-125a-5p gene was detected by RT-qPCR.The autophagy of each group was detected by MDC and Hoechst double staining,and the expressions of beclin-1 and LC3Ⅱwere detected by Western blot.The relative expressions of NF-κB/MAPK signaling pathway related genes and proteins were detected by RT-qPCR and Western blot.Results The relative expression levels of miR-125a-5p gene in the blank group,miR-NC group and miR-125a-5p group were 0.44±0.08,0.42±0.07,1.27±0.16,respectively.The relative expression level of miR-125a-5p gene in the miR-125a-5p group was higher than that in the blank group and miR-NC group,and the difference was statistically significant(P<0.05).The cell positive rates of the blank group,miR-NC group and miR-125a-5p group were(2.10±0.52)%,(2.30±0.47)%and(26.40±7.14)%,respectively.The cell positive rate of the miR-125a-5p group was higher than that of the blank group and miR-NC group,and the differences were statistically significant(P<0.05).The relative expression levels of Beclin1 and LC3II in the miR-125a-5p group was higher than that in the blank group and miR-NC group,and the differences were statistically significant(P<0.05).There was no statistically significant difference in the relative expression levels of NF-κB p65,extracellular regulatory protein kinase(ERK1/2),MAPK p38 mRNA among 3 groups(P>0.05).The ratios of NF-κB/MAPK signaling pathway phosphorylation protein to corresponding protein(NF-κB p65/NF-κB p65,p-ERK1/2/ERK1/2,p-MAPK p38/MAPK p38)in the miR-125a-5p group were higher than those in th

关 键 词：乳腺肿瘤 自噬 微小RNA-125a-5p 核因子-ΚB 有丝分裂原活化蛋白激酶 信号传导 

分 类 号：R737.9[医药卫生—肿瘤]