摘要
目的探讨PR结构域蛋白5(PRDM5)过表达对人急性髓系白血病(AML)细胞系U937迁移侵袭的影响及其机制。方法收集20例AML患者(AML组)和20例健康志愿者(正常组),采用实时荧光定量聚合酶链反应(RT-qPCR)检测其骨髓样本中微小RNA(miR)-489-3p和PRDM5 mRNA表达水平。采用脂质体转染技术将人AML细胞系U937分为7组:pcDNA3.1组、pcDNA3.1-PRDM5组、shNC组、shPRDM5组、NC组、miR-489-3p mimics组和miR-489-3p+pcDNA3.1-PRDM5组,采用划痕实验和侵袭迁移小室(Transwell小室)法分别检测各组细胞的迁移和侵袭能力;采用蛋白质免疫印迹法(Western blot)检测β-catenin和GSK3β蛋白表达水平。结果AML组受试者骨髓miR-489-3p表达水平低于正常组,PRDM5 mRNA表达水平高于正常组(P<0.05)。pcDNA3.1-PRDM5组和miR-489-3p+pcDNA3.1-PRDM5组PRDM5 mRNA、β-catenin表达水平、穿过基质胶膜的细胞数量及24 h内迁移率分别高于pcDNA3.1组、miR-489-3p mimics组,而GSK3β表达水平分别低于pcDNA3.1组、miR-489-3p mimics组(P<0.05)。shPRDM5组和miR-489-3p mimics组的PRDM5 mRNA、β-catenin表达水平、穿过基质胶膜的细胞数量及24 h内迁移率分别低于shNC组和NC组,而GSK3β蛋白表达水平分别高于shNC组和NC组(P<0.05)。结论miR-489-3p靶向PRDM5基因阻断Wnt/β-catenin信号通路,从而抑制人AML细胞系U937迁移侵袭。
Objective To explore the effect of PR domain-containing 5( PRDM5) overexpression on migration and invasion of human acute myeloid leukemia( AML) cell line U937 and its mechanism.Methods Twenty patients with AML( AML group) and 20 healthy volunteers( normal group) were collected. Real-time fluorescent quantitative polymerase chain reaction( RT-qPCR) was used to detect expression levels of microRNA( miR)-489-3 p and PRDM5 mRNA in bone marrow samples. Human AML cell line U937 were divided into 7 groups by liposome transfection technology: pc DNA3. 1 group,pc DNA3. 1-PRDM5 group,sh NC group,sh PRDM5 group,NC group,miR-489-3 p mimics group and miR-489-3 p + pc DNA3. 1-PRDM5 group. Scratch test and invasion and migration chamber( Transwell chamber) method were used to detect cell migration and invasion ability of each group respectively. Western blotting was used to detect expression levels of β-catenin and GSK3β protein. Results Expression level of miR-489-3 p in bone marrow of AML group was lower than that of normal group,and expression level of PRDM5 mRNA was higher than that of normal group( P < 0. 05). Expression levels of PRDM5 mRNA,β-catenin,number of cells passing through the matrigel membrane and migration rate within 24 h in pc DNA3. 1-PRDM5 group and miR-489-3 p + pc DNA3. 1-PRDM5 group were higher than those in pc DNA3. 1 group and miR-489-3 p mimics group respectively,expression level of GSK3β was lower than those in pc DNA3. 1 group and miR-489-3 p mimics group respectively( P < 0. 05). Expression levels of PRDM5 mRNA,β-catenin,number of cells passing through the matrigel membrane and migration rate within 24 h in sh PRDM5 group and miR-489-3 p mimics group were lower than those in sh NC group and NC group respectively,expression level of GSK3β was higher than those in sh NC group and NC group respectively( P < 0. 05). Conclusion MiR-489-3 p targets PRDM5 gene blockes the Wnt/β-catenin signal pathway,thereby inhibits the migration and invasion of human AML cell line U937.
作者
苏杰
杨小静
周雪
Su Jie;Yang Xiaojing;Zhou Xue(Department of Hematology,Jiangbei People’s Hospital,Nanjing 210048,China)
出处
《临床内科杂志》
CAS
2021年第12期836-839,共4页
Journal of Clinical Internal Medicine
基金
中石化南北公司临床医学专项科研项目(ZX2020006)。
