[1]张 璐,郭含梦,王庆义.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[J].陕西医学杂志,2024,(7):895-899.[doi:DOI:10.3969/j.issn.1000-7377.2024.07.006]
　ZHANG Lu,GUO Hanmeng,WANG Qingyi.LncRNA MEG8 regulates HMGA1 expression and the mechanism of cell proliferation and apoptosis in acute myeloid leukemia cells through miR-495-3p[J].,2024,(7):895-899.[doi:DOI:10.3969/j.issn.1000-7377.2024.07.006]
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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
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年7期
页码:
895-899
栏目:
基础研究
出版日期:
2024-07-05

文章信息/Info

Title:
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
(安徽医科大学附属阜阳医院血液科,安徽 阜阳 236000)
Author(s):
ZHANG Lu; GUO Hanmeng; WANG Qingyi
(Department of Hematology,Fuyang Hospital Affiliated to Anhui Medical University,Fuyang 236000,China)
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
Keywords:
Long non-coding RNA;  Maternal expression gene 8;  miR-495-3p;  Acute myeloid leukemia cells;  High mobility group protein 1
分类号:
R -33
DOI:
DOI:10.3969/j.issn.1000-7377.2024.07.006
文献标志码:
A
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。
Abstract:
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 HL-60 cells in si-lncRNA MEG8+miR-495-3p mimic group was higher than that in si-lncRNA MEG8 group and miR-495-3p mimic group compared with HL-60 group,si-NC group and MImea-NC group(all P<0.05).Compared with HL-60 group,si-NC group and mimic-NC group,the HMGA1 protein expression of si-lncRNA MEG8+miR-495-3p mimic group was lower than that of si-lncRNA MEG8 group and miR-495-3p mimic group(all P<0.05).Conclusion:Silencing lncRNA MEG8 may down-regulate HMGB1 through up-regulation of miR-495-3p,thereby inhibiting the malignant biological behavior of HL-60 cells,providing a new idea for exploring potential therapeutic targets for AML.

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备注/Memo

备注/Memo:
基金项目:安徽省自然科学基金资助项目(2208085MH217)
更新日期/Last Update: 2024-07-04