β-arrestin1通过miR-652-5p促进T-ALL细胞线粒体活性氧含量的研究  

β-arrestin1 Promotes the Concentration of Mitochondrial Reactive Oxygen in T-ALL Cells via MiR-652-5p

作　　者：王皓飚 刘姗[1,2,3,4] 郭维 刘海燕[2,3,4,5] 于洁 邹琳 WANG Hao-Biao;LIU Shan;GUO Wei;LIU Hai-Yan;YU Jie;ZOU Lin(Center for Clinical Molecular Medicine of Children's Hospital in Chongqing Medical University,Chongqing 400014,China;National Clinical Research Center for Child Health and Disorders,Chongqing 400014,China;Ministry of Education Key Laboratory of Child Development and Disorders,Chongqing 4000M,China;Chongqing Key Laboratory of Pediatrics,Chongqing 4000M,China;Center for Clinical Hematology,Children's Hospital in Chongqing Medical University,Chongqing 400014,China;Clinical Research Unit of Children's Hospital in Shanghai Jiaotong University,Shanghai 200062,China)

机构地区：[1]重庆医科大学附属儿童医院临床分子医学中心 [2]国家儿童健康与疾病临床医学研究中心 [3]儿童发育疾病研究教育部重点实验室 [4]儿科学重庆市重点实验室 [5]重庆医科大学附属儿童医院血液中心,重庆400014 [6]上海交通大学附属儿童医院临床研究中心,上海200062

出　　处：《中国实验血液学杂志》2021年第5期1456-1461,共6页Journal of Experimental Hematology

基　　金：国家自然科学基金(81870126,82070167,81800186)。

摘　　要：目的:探讨β-arrestin1对急性T淋巴细胞白血病(T-ALL)细胞线粒体内活性氧含量(ROS)的影响及可能作用机制。方法:构建稳定敲低β-arrestin1(Siβ1)的T-ALL细胞株Jurkat细胞。采用流式细胞术、探针法分别检测细胞及线粒体内ROS含量。microRNA芯片检测及分析和Q-PCR验证β-arrestin1与microRNA的关系。miRbase软件预测microRNA靶基因,Western blot检测microRNA靶基因表达,双荧光素酶报告基因实验验证结合。结果:成功构建Jurkat Siβ1稳定细胞株,Jurkat Siβ1整个细胞水平ROS含量略降低,且线粒体内ROS含量明显降低。microRNA芯片分析发现,多种与T-ALL相关的microRNA呈差异表达,其中miR-652-5p在Jurkat Siβ1中的表达显著升高(P<0.05 fold>2.0),且Q-PCR显示miR-652-5p在Jurkat Siβ1中上升近5倍;通过miRbase软件预测到P62基因是miR-652-5p靶基因,且其能调控线粒体功能,在miR-652-5p稳定敲低Jurkat细胞中高表达,双荧光素酶报告基因实验证实P62是miR-652-5p靶基因。结论:T-ALL中,β-arrestin1可降低miR-652-5p表达,解除对P62基因的抑制,增加细胞线粒体内ROS含量。Objective:To investigate the effect ofβ-arrestinl on the concentration of reactive oxygen species(ROS)in the mitochondria of acute T-lymphocytic leukemia(T-ALL)cells and its possible mechanisms.Methods:The stable T-ALL cell line with knocked downβ-arrestinl(Jurkat Siβ1)was constructed.Flow cytometry and probe assays were used to detect ROS content in cell and mitochondrial,respectively.The relationship betweenβ-arrestinl and microRNA was detected,analyzed and Q-PCR confirmed by microRNA microarray.The target genes of microRNA were predicated by miRbase software,identified by Western blot,and validated by Dual luciferase reporter gene.Results:Jurkat Siβ1 stable cell line was successfully constructed and it was found that ROS content was slightly reduced in Jurkat Siβ1 at the whole cell level,and the ROS content was also significantly reduced in mitochondria.MicroRNA microarray analysis revealed that multiple T-ALL related microRNAs showed differentially expressed,in which the expression of miR-652-5 p was significantly increased in Jurkat Siβ1(P<0.05 fold>2.0),and Q-PCR showed that miR-652-5 p was nearly 5-fold up-regulated in Jurkat Siβ1.miRbase predicted that the P62 gene was the target gene of miR-652-5 p which could regulates mitochondrial function.P62 protein showed highly expressed in stably knocked down miR-652-5 p in Jurkat cells.Dual luciferase reporter gene assay confirms that P62 was the target gene of miR-652-5 p.Conclusion:β-arrestinl can decreases the expression of miR-652-5 p and deregulates the translational inhibition of P62 mRNA,thus to increase ROS content in mitochondria of T-ALL cells.

关 键 词：β-arrestin1 急性T淋巴细胞白血病 活性氧 miR-652-5p 

分 类 号：R733.71[医药卫生—肿瘤] R725[医药卫生—临床医学]