miR-186对HL-60细胞增殖、迁移的调控及机制研究  

Regulation and mechanism of miR-186 on proliferation and migration of HL-60 cells

作　　者：刘洋 高长俊[2] LIU Yang;GAO Changjun(Department of Clinical Laboratory,the Second Central Hospital of Baoding in Hebei Province,Baoding,Hebei 072650,China;Department of Pediatric Hematology,Tangshan Maternal and Child Health Hospital in Hebei Province,Tangshan,Hebei 063000,China)

机构地区：[1]河北省保定市第二中心医院检验科,河北保定072650 [2]河北省唐山市妇幼保健院小儿血液科,河北唐山063000

出　　处：《检验医学与临床》2024年第5期596-602,共7页Laboratory Medicine and Clinic

基　　金：国家卫生健康委员会“十四五”规划全国重点课题(YYWS4359)。

摘　　要：目的 探讨过表达微小RNA-186(miR-186)对人原髓细胞白血病HL-60细胞上皮间质转化(EMT)、增殖、迁移及磷脂酰肌醇3-激酶(PI3K)/丝苏氨酸蛋白激酶(AKT)信号通路的调控作用。方法 选取体外培养HL-60细胞,按照不同的转染方式分为对照组、mimic NC组、miR-186组、miR-186+抑制剂组、miR-186+激活剂组。采用实时荧光定量聚合酶链反应、5-乙炔基-2′脱氧尿嘧啶核苷(EdU)、Transwell小室法及蛋白免疫印迹法检测miR-186相对表达水平、增殖率、迁移率、EMT相关因子及其信使RNA(mRNA)相对表达水平并进行比较、分析。结果 与mimic NC组相比,miR-186组HL-60细胞miR-186相对表达水平、E-钙黏蛋白(E-cadherin)及其mRNA表达水平升高,差异均有统计学意义(P<0.05),而增殖率、迁移率、磷酸化(p)-PI3K、p-AKT、N-钙黏蛋白(N-cadherin)、波形蛋白(Vimentin)、纤维粘连蛋白(FN)及其mRNA表达水平降低,差异均有统计学意义(P<0.05)。与miR-186组相比,miR-186+抑制剂组HL-60细胞E-cadherin及其mRNA表达水平升高,差异均有统计学意义(P<0.05),而增殖率、迁移率、p-PI3K、p-AKT、N-cadherin、Vimentin、FN及其mRNA表达水平降低,差异均有统计学意义(P<0.05);与miR-186组相比,miR-186+激活剂组HL-60细胞E-cadherin及其mRNA表达水平下降,差异均有统计学意义(P<0.05),而增殖率、迁移率、p-PI3K、p-AKT、N-cadherin、Vimentin、FN及其mRNA表达水平升高,差异均有统计学意义(P<0.05)。结论 过表达miR-186通过阻遏PI3K/AKT信号通路抑制HL-60细胞EMT,进而抑制HL-60细胞增殖和迁移。Objective To explore the regulatory effects of overexpression of microRNA-186(miR-186) on epithelial mesenchymal transformation(EMT),proliferation,migration and phosphatidylinoinositide 3-kinase(PI3K)/seronine protein kinase(AKT) signaling pathway of human promyelocytic leukemia HL-60 cells.Methods HL-60 cells cultured in vitro were divided into control group,mimic NC group,miR-186 group,miR-186+inhibitor group and miR-186+activator group according to different transfection contents.Real-time fluorescence quantitative PCR,5-acetyney-2′deoxyuracil nucleoside(EdU),Transwell and Western blotting were used to analyze the miR-186 relative expression level,proliferation rate,mobility and EMT-related factors relative expression levels.Results Compared with mimic NC group,the relative expression levels of miR-186,E-cadherin and its mRNA in HL-60 cells of miR-186 group were increased,and the differences were statistically significant(P<0.05).Proliferation rate,mobility,phosphorylation(p)-PI3K,p-AKT,N-cadherin,Vimentin,fibronectin(FN) and their massenger RNA(mRNA) expression levels were decreased,and the differences were statistically significant(P<0.05).Compared with miR-186 group,the expression levels of E-cadherin and its mRNA of HL-60 cells in miR-186+ inhibitor group was increased,and the differences were statistically significant(P<0.05).Proliferation rate,mobility,p-PI3K,p-AKT,N-cadherin,Vimentin,FN and their mRNA expression levels were decreased,and the differences were statistically significant(P<0.05).Compared with miR-186 group,the expression levels of E-cadherin and its mRNA of HL-60 cells in miR-186+ activator group decreased,and the differences were statistically significant(P<0.05).Proliferation rate,mobility,p-PI3K,p-AKT,N-cadherin,Vimentin,FN and their mRNA expression levels were increased,and the differences were statistically significant(P<0.05).Conclusion Overexpression of miR-186 inhibits EMT in HL-60 cells by repressing PI3K/AKT signaling pathway,thereby inhibiting cell proliferation and migration.

关 键 词：微小RNA-186 人原髓细胞白血病 HL-60细胞 磷脂酰肌醇3-激酶/丝苏氨酸蛋白激酶信号通路 增殖 迁移 

分 类 号：R733.71[医药卫生—肿瘤]