摘要
目的长链非编码RNA人类白细胞抗原复合体18(lncRNA HCG18)调对急性髓系白血病(AML)的作用机制尚不清楚。探讨lncRNA HCG18控微小RNA-324-5p(miR-324-5p)/胶质瘤相关基因同源蛋白1(GLI1)轴对AML细胞增殖、迁移与侵袭的影响。方法选择2017年1月-2021年3月在本院血液科住院治疗的AML患者35例作为研究对象(AML组),另选取同期在本院治疗并进行骨髓穿刺检查的非血液系统肿瘤性疾病患者30例作为对照组,实时荧光定量聚合酶链反应(qRT-PCR)检测所有入选人员骨髓单个核细胞中lncRNA HCG18与miR-324-5p的表达。体外培养AML细胞HL-60,将细胞分为对照组、si-NC组、lncRNA HCG18 siRNA组、miR-NC组、miR-324-5p mimics组、miR-324-5p mimics+GLI1 NC组、miR-324-5p mimics+GLI1组、lncRNA HCG18 siRNA+NC inhibitor组、lncRNA HCG18 siRNA+miR-324-5p inhibitor组,分别检测检测HL-60细胞中lncRNA HCG18、miR-324-5p表达、HL-60细胞增殖、迁移与侵袭及GLI1、细胞周期蛋白D1(Cyclin D1)、基质金属蛋白酶(MMP)-2和MMP-9蛋白表达。双荧光素酶报告系统分析miR-324-5p与lncRNA HCG18、GLI1的靶向关系。结果与对照组比较,AML组患者骨髓单个核细胞中lncRNA HCG18表达水平(2.85±0.32)较对照组(1.03±0.11)显著升高(P<0.01),而miR-324-5p表达水平(0.47±0.05)较对照组(1.05±0.09)显著降低(P<0.01);沉默lncRNA HCG18表达或过表达miR-324-5p后HL-60细胞增殖、细胞迁移与侵袭数、Cyclin D1、MMP-2、MMP-9蛋白表达显著降低(P<0.05);双荧光素酶报告基因实验显示,miR-324-5p与lncRNA HCG18、GLI1均有靶向关系;抑制miR-324-5p表达可逆转沉默lncRNA HCG18表达对HL-60细胞增殖、迁移与侵袭的抑制作用;GLI1过表达可逆转过表达miR-324-5p对HL-60细胞增殖、迁移与侵袭的抑制作用。结论lncRNA HCG18在AML细胞中高表达,抑制lncRNA HCG18表达可通过调控miR-324-5p/GLI1轴抑制AML细胞增殖、迁移与侵袭。
Objective The mechanism of long noncoding RNA human leukocyte antigen complex 18(lncRNA HCG18)on acute myeloid leukemia(AML)has not been well understood.The paper was to investigate how lncRNA HCG18 impacts the proliferation,migration and invasion of AML cells by regulating microRNA-324-5p(miR-324-5p)/glioma-related gene homologous protein 1(GLI1)axis.Methods Thirty-five AML patients who were hospitalized at the Department of Hematology in our hospital from January 2017 to March 2021 were registered(AML group).Separately,30 patients with non-hematological tumor diseases undergoing bone marrow puncture in our hospital during the same period were selected as the control group.Real-time fluorescent quantitative polymerase chain reaction(qRT-PCR)was employed to detect the expression of lncRNA HCG18 and miR-324-5p in bone marrow mononuclear cells of all selected personnel.AML cells HL-60 were cultured in vitro and divided into control group,si-NC group,lncRNA HCG18 siRNA group,miR-NC group,miR-324-5p mimics group,miR-324-5p mimics+GLI1 NC group,miR-324-5p mimics+GLI1 group,lncRNA HCG18 siRNA+NC inhibitor group,lncRNA HCG18 siRNA+miR-324-5p inhibitor group,then the expression of lncRNA HCG18,miR-324-5p of HL-60 cells,and HL-60 cells proliferation,migration and invasion,GLI1,Cyclin D1,matrix metalloproteinase(MMP)-2 and MMP-9 were examined respectively.The dual luciferase reporter system was used to analyze the targeting relationship between miR-324-5p and lncRNA HCG18,GLI1.Results The expression level of lncRNA HCG18(2.85±0.32)in bone marrow mononuclear cells of the AML group was significantly increased(P<0.01)when compared with the control group(1.03±0.11),yet the expression level of miR-324-5p was significantly decreased(P<0.01).The numbers of proliferation,cell migration and invasion of HL-60 cells,Cyclin D1,MMP-2,and MMP-9 protein expression were significantly reduced(P<0.05)after silencing lncRNA HCG18 expression or overexpressing miR-324-5p.Dual luciferase reporter gene experiment indicated that miR-324-5p had a targeting relationship with lncRNA HCG18 and GLI1.Moreover,inhibition of miR-324-5p expression,as well as the overexpression of GLI1,could reverse the inhibitory effects of silencing lncRNA HCG18 expression on the proliferation,migration and invasion of HL-60 cells.Conclusion LncRNA HCG18 is highly expressed in AML cells.Additionally,inhibiting the expression of lncRNA HCG18 can inhibit the proliferation,migration and invasion of AML cells by regulating the miR-324-5p/GLI1 axis.
作者
刘敏
雷荟融
成延娟
唐元艳
黄知平
LIU Min;LEI Hui-rong;CHENG Yan-juan;TANG Yuan-yan;HUANG Zhi-ping(Department of Hematology,Jingzhou Hospital Affiliated to Yangtze University,Jingzhou 434020,Hubei,China)
出处
《医学研究生学报》
CAS
北大核心
2022年第5期485-492,共8页
Journal of Medical Postgraduates
基金
湖北省卫生健康委员会科研项目(WJ2019 M081)。
作者简介
刘敏,医学硕士;通信作者:黄知平,E-mail:191060635@qq.com。
