miR-760/CPEB2轴对急性淋巴细胞白血病细胞凋亡和增殖的影响  

Effect of miR-760/CPEB2 axis on apoptosis and proliferation in acute lymphoblastic leukemia

作　　者：刘述川 郭强 张宇晶[1] 林婧祎 张颖[1] LIU Shuchuan;GUO Qiang;ZHANG Yujing;LIN Jingyi;ZHANG Ying(Department of Hemotology,the 4th Affiliated Hospital of Harbin Medical University,Harbin 150001,Heilongjiang,China)

机构地区：[1]哈尔滨医科大学附属第四医院血液科,哈尔滨150001

出　　处：《医学研究与战创伤救治》2024年第4期346-351,共6页Journal of Medical Research & Combat Trauma Care

基　　金：哈尔滨医科大学附属第四医院特别资助科研项目(HYDSYTB202225)。

摘　　要：目的探究miR-760对急性淋巴细胞白血病(ALL)凋亡和增殖的影响和机制。方法采用实时荧光定量PCR(qRT-PCR)检测miR-760和细胞质多腺苷酸化元件结合蛋白2(CPEB2)相对表达。Ball-1细胞分为不同组:miR-NC组(转染miR-NC)、miR-760mimic组(转染miR-760 mimic)、si-NC组(转染si-NC)、si-CPEB2组(转染si-CPEB2)、oe-NC组(转染oe-NC)、oe-CPEB2组(转染oe-CPEB2)、miR-760 mimic+oe-NC组(共转染miR-760 mimic+oe-NC)和miR-760 mimic+oe-CPEB2组(共转染miR-760 mimic+oe-CPEB2)。流式细胞术检测细胞凋亡,Western blot检测Bax、c-caspase-3、t-caspase-3、Ki67、PCNA和CPEB2相对表达量。双荧光素酶报告实验验证miR-760和CPEB2靶向结合。结果与人B淋巴母细胞HMy2.CIR比较,ALL细胞系Ball-1、Reh和JurkatE6中miR-760水平表达下降,CPEB2 mRNA和蛋白水平表达上升(P<0.05)。与miR-NC组比较,miR-760 mimic组细胞凋亡率、Bax和c-caspase-3水平表达上升,而Ki67和PCNA水平表达下降(P<0.05)。生物信息学方法预测miR-760靶向调控CPEB2,双荧光素酶报告实验验证CPEB2是miR-760的靶标。与si-NC组比较,si-CPEB2组细胞凋亡率、Bax和c-caspase-3水平表达上升,而Ki67、PCNA水平表达下降(P<0.05)。上调CPEB2表达能逆转miR-760过表达对细胞凋亡率、Bax、ccaspase-3的促进和对Ki67、PCNA的抑制(P<0.05)。结论miR-760靶向调节CPEB2,诱导细胞凋亡、抑制细胞增殖,为ALL治疗提供新选择。Objective This study was aimed to explore the effect and mechanism of miR-760 on apoptosis and proliferation of lymphoblasts in acute lymphoblastic leukemia(ALL).Methods Real-time fluorescence quantitative PCR(qRT-PCR)was used to detect the relative expression of miR-760 and cytoplasmic polyadenylation element binding protein 2(CPEB2).Human B-lymphocyte acute leukemia cells Ball-1 were transfected with miR-NC,miR-760 mimic,si-NC,si-CPEB2,oe-NC,oe-CPEB2,miR-760 mimic+oe-NC and miR-760 mimic+oe-CPEB2,respectively=.Cell apoptosis was examined by flow cytometry.Relative expression of Bax,c-caspase-3,t-caspase-3,Ki67,PCNA and CPEB2 was detected by Western blotting.The targeting relationship of miR-760 and CPEB2 was verified by luciferase report experiment.Results Compared with human B lymphoblasts HMy2.CIR cells,miR-760 expression was decreased in ALL cell lines Ball-1,Reh and JurkatE6,while CPEB2 mRNA and protein expression was increased(P<0.05).Compared with miR-NC group,apoptosis rate and protein expression of Bax in miR-760 mimic group was increased,while Ki67 and PCNA expression was decreased(P<0.05).Bioinformatics methods predicted that miR-760 targeted CPEB2.Dual-luciferase reporter assay verified that CPEB2 was the target of miR-760.Compared with si-NC group,the apoptosis rate,Bax and c-caspase-3 expression were increased,while protein expression of Ki67 and PCNA was decreased in si-CPEB2 group(P<0.05).Up-regulation of CPEB2 reversed the promotion of miR-760 overexpression on cell apoptosis rate,Bax and c-caspase-3 expression,as well as its inhibition on protein expression of Ki67 and PCNA(P<0.05).Conclusion MiR-760 targets CPEB2 to activate cell apoptosis and inhibit cell proliferation,which provides a new option for ALL treatment.

关 键 词：miR-760 急性淋巴细胞白血病 凋亡 增殖 细胞质多腺苷酸化元件结合蛋白2 

分 类 号：R733.71[医药卫生—肿瘤]