[1]敖会芳,黄 华,王红权,等.长链非编码RNA RP11-641D5.1通过靶向微小RNA-486-5p调控急性髓系白血病细胞增殖、细胞周期和免疫逃逸实验研究[J].陕西医学杂志,2024,(3):297-302.[doi:DOI:10.3969/j.issn.1000-7377.2024.03.002]
　AO Huifang,HUANG Hua,WANG Hongquan,et al.Long non-coding RNA RP11-641D5.1 regulates cell proliferation,cell cycle and immune escape in acute myeloid leukemia cells by targeting microRNA-486-5P[J].,2024,(3):297-302.[doi:DOI:10.3969/j.issn.1000-7377.2024.03.002]
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长链非编码RNA RP11-641D5.1通过靶向微小RNA-486-5p调控急性髓系白血病细胞增殖、细胞周期和免疫逃逸实验研究
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《陕西医学杂志》[ISSN:1000-7377/CN:61-1281/TN]

卷:
期数:
2024年3期
页码:
297-302
栏目:
基础研究
出版日期:
2024-03-05

文章信息/Info

Title:
Long non-coding RNA RP11-641D5.1 regulates cell proliferation,cell cycle and immune escape in acute myeloid leukemia cells by targeting microRNA-486-5P
作者:
敖会芳1; 黄 华1; 王红权1; 刘 俊2; 郭春梅3; 姚 云1
(1.荆楚理工学院附属中心医院 荆门市人民医院血液内科,湖北 荆门 448000; 2.荆楚理工学院附属中心医院 荆门市人民医院感染性疾病科,湖北 荆门 448000; 3.大连医科大学附属第一医院肿瘤科,辽宁 大连 116011)
Author(s):
AO Huifang; HUANG Hua; WANG Hongquan; LIU Jun; GUO Chunmei; YAO Yun
(Department of Hematology,Affiliated Central Hospital of Jingchu University of Technology,Jingmen 448000,China)
关键词:
急性髓系白血病;  RP11-641D5.1;  微小RNA-486-5p;  细胞增殖;  细胞周期;  免疫逃逸
Keywords:
Acute myeloid leukemia;  RP11-641D5.1;  miR-486-5p;  Cell proliferation;  Cell cycle;  Immune escape
分类号:
R 73-3
DOI:
DOI:10.3969/j.issn.1000-7377.2024.03.002
文献标志码:
A
摘要:
目的:探讨长链非编码RNA(lncRNA)RP11-641D5.1对急性髓系白血病细胞增殖、细胞周期及免疫逃逸的调控作用及机制。方法:采用实时定量PCR(RT-qPCR)分析RP11-641D5.1在急性髓系白血病细胞株(HL-60、SKM-1、THP-1、KG-1、NB4)和人骨髓基质细胞HS-5中的表达量。分别将阴性质粒(NC组)和RP11-641D5.1质粒(RP11-641D5.1组)转染至SKM-1细胞。采用集落形成实验和流式细胞术检测转染后各组SKM-1细胞增殖和细胞周期。采用酶联免疫吸附实验检测两组细胞干扰素-γ(IFN-γ)、肿瘤坏死因子-α(TNF-α)、白细胞介素-2(IL-2)的表达量。采用LncCeRBase软件分析RP11-641D5.1与miR-486-5p的靶向关系,并采用双荧光素酶报告实验验证。采用RT-qPCR检测RP11-641D5.1对微小RNA(miR)-486-5p表达的调控作用。采用蛋白质印迹(Western blot)检测酪氨酸激酶(JAK)-信号转导和转录激活因子3(STAT3)信号通路中磷酸化酪氨酸激酶(p-JAK)、磷酸化转录激活因子(p-STAT)、细胞因子信号转导抑制因子3(SOCS3)、核内原癌基因c-myc蛋白的表达。结果:相对于HS-5细胞,急性髓系白血病细胞株HL-60、SKM-1、THP-1、KG-1、NB4中RP11-641D5.1表达较低,且SKM-1细胞中表达最低(均P<0.05),故后续采用SKM-1细胞进行实验。与NC组比较,过表达RP11-641D5.1能够抑制SKM-1细胞增殖和细胞周期进展,促进细胞因子IFN-γ、TNF-α、IL-2的表达(均P<0.05)。RP11-641D5.1能够靶向结合miR-486-5p(P<0.05)。与NC组比较,RP11-641D5.1组细胞中miR-486-5p表达下调,JAK-STAT3信号通路相关蛋白表达水平降低(均P<0.05)。结论:RP11-641D5.1在急性髓系白血病中表达水平降低,可能通过下调miR-486-5p表达抑制急性髓系白血病细胞增殖、细胞周期进展和免疫逃逸。
Abstract:
Objective:To investigate the regulatory effect and mechanism of long non-coding RNA(lncRNA)RP11-641D5.1 on the proliferation,cell cycle and immune escape of acute myeloid leukemia cells.Methods:The expression of RP11-641D5.1 in acute myeloid leukemia cell lines(HL-60,SKM-1,THP-1,KG-1,NB4)and human bone marrow stromal cells HS-5 was analyzed by RT-qPCR.The negative plasmid(NC group)and RP11-641D5.1 plasmid(RP11-641D5.1 group)were transfected into SKM-1 cells respectively.Colony formation assay and flow cytometry were used to detect the proliferation and cell cycle of SKM-1 cells in each group after transfection.The expression levels of IFN-γ,TNF-α,and IL-2 in the two groups were detected by enzyme-linked immunosorbent assay.The targeting relationship between RP11-641D5.1 and miR-486-5p was analyzed by LncCeRBase software and verified by dual luciferase reporter assay.The regulatory effect of RP11-641D5.1 on the expression of miR-486-5p was detected by qRT-PCR.Western blot was used to detect the expression levels of janus activated kinase(JAK)-signal transducer and activator of transcription 3(STAT3)signaling pathway proteins p-JAK,p-STAT,suppressor of cytokine signaling 3(SOCS3)and c-myc.Results:Compared with HS-5 cells,the expression of RP11-641D5.1 in acute myeloid leukemia cell lines HL-60,SKM-1,THP-1,KG-1 and NB4 was lower,and the expression of RP11-641D5.1 was the lowest in SKM-1 cells(all P<0.05).Therefore,SKM-1 cells were used for subsequent experiments.Compared with the NC group,overexpression of RP11-641D5.1 could inhibit the proliferation of SKM-1 cells and cell cycle progression,and promote the the expressions of cytokine IFN-γ,TNF-α,IL-2(all P<0.05).RP11-641D5.1 could target miR-486-5p(P<0.05).Compared with NC group,the expression of miR-486-5p in RP11-641D5.1 group was down-regulated,and the expression level of JAK-STAT3 signaling pathway related proteins was decreased(all P<0.05).Conclusion:The expression level of RP11-641D5.1 is decreased in acute myeloid leukemia,which may inhibit the proliferation,cell cycle progression and immune escape of acute myeloid leukemia cells by down-regulating the expression of miR-486-5p.

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备注/Memo

备注/Memo:
基金项目:国家自然科学基金资助项目(31900517)
更新日期/Last Update: 2024-03-05