miR-4286调控FOXO4对肺癌细胞增殖和转移的影响  

Effects of miR 4286 on the proliferation and metastasis of lung cancer cells by regulating FOXO4

作　　者：何智慧 黄丽 苏菊红 刘峰[1] HE Zhihui;HUANG Li;SU Juhong(Department of ICU,Chongqing Seventh People’s Hospital,Chongqing 450000,China)

机构地区：[1]重庆市第七人民医院重症医学科,450000 [2]西南医科大学基础医学院解剖与组胚教研室,四川省泸州市646000

出　　处：《河北医药》2021年第10期1460-1464,共5页Hebei Medical Journal

摘　　要：目的探讨微小RNA-4286(miR-4286)对肺癌细胞增殖、迁移和侵袭的影响。方法实时荧光定量PCR(RT-qPCR)检测人肺癌细胞系A549和人正常支气管上皮细胞系HBE中miR-4286和FOXO4 mRNA的表达水平。转染miR-4286抑制剂或FOXO4过表达载体至肺癌细胞A549,构建miR-4286表达抑制或过表达FOXO4的肺癌细胞A549。四甲基噻唑蓝染色法(MTT)检测细胞存活率、Transwell法检测细胞的迁移和侵袭能力、蛋白印迹法(Western blot)检测细胞周期蛋白1(CyclinD1)、P21、基质金属蛋白酶-2(MMP-2)、基质金属蛋白酶-9(MMP-9)蛋白表达。双荧光素酶报告基因实验验证miR-4286与FOXO4靶向关系。结果与正常支气管上皮细胞组比较,肺癌细胞中miR-4286表达显著升高(P<0.05),FOXO4 mRNA表达显著降低(P<0.05)。抑制miR-4286表达或过表达FOXO4后,肺癌细胞存活率、迁移和侵袭细胞数、CyclinD1蛋白、MMP-2蛋白、MMP-9蛋白表达显著降低(P<0.05),FOXO4 mRNA、P21蛋白表达显著升高(P<0.05)。miR-4286在肺癌细胞中靶向负调控FOXO4表达。抑制FOXO4表达可逆转干扰miR-4286对肺癌细胞增殖、迁移和侵袭的影响。结论抑制miR-4286表达通过靶向上调FOXO4表达抑制肺癌细胞的增殖、迁移和侵袭。Objective To investigate the effects of microRNA 4286(miR-4286)on the proliferation,migration and invasion of lung cancer cells.Methods Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the mRNA expression levels of miR 4286 and FOXO4 in human lung cancer cell line A549 and human normal bronchial epithelial cell line HBE.Lung cancer cell A549 was transfected with miR 4286 inhibitor or FOXO4 overexpression vector,and lung cancer cell A549 was constructed with miR 4286 expression inhibited or overexpressed FOXO4.The methylthiazole tetrazolium(MTT)assay was used to detect cell survival rate,and Transwell method was used to detect cell migration and invasion ability.Western blot method was used to detect the protein expression of CyclinD1,P21,matrix metalloproteinase-2(MMP-2),matrix metalloproteinase-9(MMP-9).The dual luciferase reporter gene experiment was used to verify the targeting relationship between miR-4286 and FOXO4.Results As compared with those in normal bronchial epithelial cell,the expression levels of miR-4286 in lung cancer cells were significantly increased(P<0.05),while the mRNA expression levels of FOXO4 were significantly decreased(P<0.05).After inhibiting the expression of miR-4286 or overexpressing FOXO4,the survival rate of lung cancer cells,the number of cells that migrated and invaded,and the expression levels of CyclinD1 protein,MMP-2 protein,and MMP-9 protein were significantly decreased(P<0.05),while the expression levels of FOXO4 mRNA and P21 protein were significantly increased(P<0.05).The miR-4286 targeted and negatively regulated FOXO4 expression in lung cancer cells.To inhibit FOXO4 expression could reverse the effects of miR-4286 on the proliferation,migration and invasion of lung cancer cells.Conclusion To inhibit miR-4286 expression can suppress the proliferation,migration and invasion of lung cancer cells by targeting up-regulation of FOXO4 expression.

关 键 词：miR-4286 FOXO4 肺癌 增殖 迁移 侵袭 

分 类 号：R734.2[医药卫生—肿瘤]