[1]杨瑞,宁婧,景丽娟,等.miRNA-495靶向STYX调节MLL-AF9的急性髓系白血病细胞的增殖和凋亡[J].陆军军医大学学报(原第三军医大学学报),2021,43(22):2407-2413.
 YANG Rui,NING Jing,JING Lijuan,et al.MiRNA-495 regulates proliferation and apoptosis of acute myeloid leukemia cells with MLL gene rearrangement by targeting STYX[J].J Amry Med Univ (J Third Mil Med Univ),2021,43(22):2407-2413.
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miRNA-495靶向STYX调节MLL-AF9的急性髓系白血病细胞的增殖和凋亡(/HTML )
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陆军军医大学学报(原第三军医大学学报)[ISSN:1000-5404/CN:51-1095/R]

卷:
43卷
期数:
2021年第22期
页码:
2407-2413
栏目:
基础医学
出版日期:
2021-11-30

文章信息/Info

Title:
MiRNA-495 regulates proliferation and apoptosis of acute myeloid leukemia cells with MLL gene rearrangement by targeting STYX
作者:
杨瑞宁婧景丽娟王海楠崔丽娟
宁夏医科大学总医院:血液科,医学实验中心
Author(s):
YANG Rui NING Jing JING Lijuan WANG Hainan CUI Lijuan
Department of Hematology, 2Medical Laboratory Center, General Hospital of Ningxia Medical University, Yinchuan, Ningxia Hui Autonomous Region, 750004, China
 
关键词:
急性髓系白血病MLL-AF9miRNA-495STYX基因细胞增殖
Keywords:
acute myeloid leukemia MLL-AF9 miRNA-495 serine/threonine/tyrosine interacting protein cell proliferation
分类号:
R394.3; R730.23; R733.71
文献标志码:
A
摘要:

目的探究miRNA-495对混合谱系白血病基因(mixed lineage leukemia, MLL)基因重排的急性髓系白血病(acute myeloid leukemia,AML)增殖和凋亡的影响。方法收集2017年8月至2019年5月于宁夏医科大学总医院血液科就医确诊为AML的10例MLL-AF9白血病患者[男性6例,女性4例,年龄(39.37±2.71)岁];10例急性髓系白血病患者[男性5例,女性5例,年龄(41.12±1.92)岁];同时间段于外科门诊就医患者10例[男性6例,女性4例,年龄(42.31±3.21)岁]。根据是否转染miRNA-495 mimic分组为miRNA-495组和Control组,通过RT-qPCR检测各组miRNA-495的表达;CCK-8检测各组转染THP-1细胞的增殖情况;RT-qPCR和Western blot检测各组转染THP-1细胞增殖相关因子(Ki-67和PCNA)和细胞凋亡相关因子(Bcl-2、Bax和Caspase-3)的表达;TargetScan预测miRNA-495的靶基因并用双荧光素酶报告实验验证;RT-qPCR和Western blot检测miRNA-495组(转染miRNA-495)、Control组、丝氨酸/苏氨酸/酪氨酸相互作用蛋白(Serine/threonine/tyrosine interacting protein,STYX)组(转染STYX)和miRNA-495+STYX组(转染miRNA-495和STYX)中STYX的表达。结果miRNA-495在MLL-AF9的AML患者中表达下调[MLL-AF9患者相对表达量(0.25±0.08),95%CI: -0.86~-0.65,P<0.01,P<0.001];miRNA-495可以抑制THP-1细胞增殖(P<0.05,P<0.01,P<0.001);miRNA-495可以诱导THP-1细胞的凋亡(P<0.05,P<0.001);miRNA-495靶向结合STYX(P<0.01,P<0.001);STYX过表达可以逆转miRNA-495对THP-1细胞增殖和凋亡的作用(P<0.05,P<0.01,P<0.001)。结论miRNA-495通过靶向STYX调控MLL基因重排的急性髓系白血病细胞的增殖和凋亡。

Abstract:

ObjectiveTo investigate the effect of miRNA-495 on the proliferation and apoptosis of acute myeloid leukemia (AML) cells with mixed lineage leukemia (MLL) gene rearrangement.  MethodsTen patients diagnosed with AML with MLL-AF9 fusion gene positive (6 males, 4 females, mean age of 39.37±2.71 years), and 10 cases of AML (5 males, 5 females, mean age of 41.12±1.92 years) admitted in our hospital from August 2017 to May 2019 were enrolled in this study, and another 10 patients (6 males, 4 females, mean age of 42.31±3.21 years) who attended the surgical outpatient department during the same period were also collected as the control. THP-1 cells (containing MLL-AF9 fusion gene) were divided into miRNA-495 and control groups based on transfection with miRNA-495 or not, and the expression of miRNA-495 in each group was detected by RT-qPCR; CCK-8 assay was used to determine the proliferation of transfected THP-1 cells in each group; RT-qPCR and Western blotting were performed to measure the expression of proliferation-related factors (Ki-67 and PCNA) and apoptosis-related factors (Bcl-2, Bax and Caspase-3). The target genes of miRNA-495 were predicted by TargetScan and then verified by dual-luciferase reporter assay. Finally, RT-qPCR and Western blotting were adopted to detect the expression of serine/threonine/tyrosine interacting protein (STYX) in groups as follows: miRNA-495 group, control group, STYX group (transfected with STYX) and miRNA-495+STYX group (transfected with miRNA-495 and STYX). ResultsMiRNA-495 expression was down-regulated in AML patients with MLL-AF9 fusion gene, with relative expression of 0.25±0.08 (95%CI=-0.86~-0.65, P<0.001); miRNA-495 inhibited cell proliferation (P<0.001) and induced apoptosis of THP-1 cells, and it targetedly bound to STYX (P<0.001). In addition, STYX overexpression reversed the effects of miRNA-495 on THP-1 cell proliferation and apoptosis. ConclusionMiRNA-495 regulates the proliferation and apoptosis of AML cells with MLL gene rearrangement by targeting STYX.

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更新日期/Last Update: 2021-11-23