长链非编码RNA母系表达基因8通过微小RNA-495-3p调控急性髓性白血病细胞高迁移率族蛋白A1表达及对细胞增殖、凋亡的作用机制研究  

LncRNA MEG8 regulates HMGA1 expression and the mechanism of cell proliferation and apoptosis in acute myeloid leukemia cells through miR-495-3p

作　　者：张璐 郭含梦 王庆义 ZHANG Lu;GUO Hanmeng;WANG Qingyi(Department of Hematology,Fuyang Hospital Affiliated to Anhui Medical University,Fuyang 236000,China)

机构地区：[1]安徽医科大学附属阜阳医院血液科,安徽阜阳236000

出　　处：《陕西医学杂志》2024年第7期895-899,共5页Shaanxi Medical Journal

基　　金：安徽省自然科学基金资助项目(2208085MH217)。

摘　　要：目的:探讨长链非编码RNA(lncRNA)母系表达基因8(MEG8)通过微小RNA-495-3p(miR-495-3p)调控急性髓性白血病细胞(AML)高迁移率族蛋白A1(HMGA1)表达及对细胞增殖、凋亡的机制。方法:选取50例经确诊为AML患者的骨髓活检标本与52例健康骨髓捐赠者骨髓标本,常规培养人AML细胞株HL-60,采用实时荧光定量PCR(RT-qPCR)法骨髓单个核细胞中lncRNA MEG8 mRNA表达。将HL-60细胞分为六组,即HL-60组(HL-60细胞)、si-NC组(转染si-NC)、si-lncRNA MEG8组(转染si-lncRNA MEG8)、mimic-NC组(转染mimic-NC)、miR-495-3p mimic组(转染miR-495-3p mimic)、si-lncRNA MEG8+miR-495-3p mimic组(转染si-lncRNA MEG8+miR-495-3p mimic),CCK-8法检测培养24、48、72 h各组细胞增殖能力,流式细胞术检测细胞凋亡,Western blot法检测细胞中HMGA1表达,双荧光素酶报告基因实验验证lncRNA MEG8、miR-495-3p、HMGA1之间的关系。结果:与对照组相比,观察组lncRNA MEG8 mRNA表达升高(P<0.05)。与miR NC+MEG8 WT组比较,miR-495-3p mimic+MEG8 WT组荧光素酶活性下降(P<0.05),与miR NC+HMGA1 WT组比较,miR-495-3p mimic+HMGA1 WT组荧光素酶活性下降(P<0.05)。与HL-60组、si-NC组、mimic-NC组比较,si-lncRNA MEG8组、miR-495-3p mimic组和si-lncRNA MEG8+miR-495-3p mimic组24、48 h细胞增殖能力均显著下降,si-lncRNA MEG8+miR-495-3p mimic组细胞增殖率最低(P<0.05)。与HL-60组、si-NC组和mimic-NC组比较,si-lncRNA MEG8+miR-495-3p mimic组HL-60细胞凋亡率高于si-lncRNA MEG8组和miR-495-3p mimic组(均P<0.05)。与HL-60组、si-NC组和mimic-NC组比较,si-lncRNA MEG8+miR-495-3p mimic组HMGA1蛋白表达低于si-lncRNA MEG8组和miR-495-3p mimic组(均P<0.05)。结论:沉默lncRNA MEG8可能通过上调miR-495-3p来下调HMGB1,来抑制HL-60细胞的恶性生物学行为,可为探索AML潜在治疗靶点提供了新思路。Objective:To investigate the mechanism of long non-coding RNA(lncRNA)maternal expression gene 8(MEG8)regulating the expression of high mobility group protein 1(HMGA1)through miR-495-3p in acute myeloid leukemia(AML)cells,cell proliferation and apoptosis.Methods:Bone marrow biopsy samples from 50 patients diagnosed with AML and bone marrow samples from 52 healthy bone marrow donors were selected.Human AML cell line HL-60 was cultured routinely,and lncRNA MEG8 mRNA was expressed in bone marrow mononuclear cells by RT-qPCR method.The HL-60 cells were divided into six groups.That is,HL-60 group(HL-60 cells),si-NC group(transfected si-NC),si-lncRNA MEG8 group(transfected si-lncRNA MEG8),mimeo-NC group(transfected Mimeo-NC),miR-495-3p mimic group(transfected miR-495-3p mimic group)and si-lncRNA MEG8+miR-495-3p mimic group(transfected with si-lncRNA MEG8+miR-495-3p mimic group),the cell proliferation ability of each group was detected by CCK-8 method after 24,48,and 72 hours.Cell apoptosis was detected by flow cytometry.Western blot was used to detect HMGA1 expression in cells.Dual luciferase reporter gene assay verified the relationship between lncRNA MEG8,miR-495-3p and HMGA1.Results:Compared with control group,lncRNA MEG8 mRNA expression in observation group was increased(P<0.05).The luciferase activity of miR-495-3p mimic+MEG8 WT co-transfection group decreased compared with that of miR-NC+MEG8 WT co-transfection group(P<0.05).Compared with that of miR-NC+HMGA1 WT co-transfection group,the luciferase activity of miR-495-3p mimic+MEG8 WT co-transfection group decreased significantly(P<0.05).The luciferase activity of miR-495-3p mimic+HMGA1 WT co-transfected group decreased(P<0.05).Cell proliferation ability of si-lncRNA MEG8 group,miR-495-3p mimic group and si-lncRNA MEG8+miR-495-3p mimic group decreased significantly at 24 and 48 hours compared with HL-60 group,si-NC group and mimic-NC group(all P<0.05).The cell proliferation rate of si-lncRNA MEG8+miR-495-3p mimic group was the lowest(P<0.05).The apoptosis rate of

关 键 词：长链非编码RNA 母系表达基因8 微小RNA-495-3p 急性髓性白血病细胞 高迁移率族蛋白1 

分 类 号：R-33[医药卫生]