lncRNA CYTOR/miR-193b-3p/HMGB1轴调控T细胞急性淋巴细胞白血病细胞MOLT-4增殖与凋亡的作用研究  

Role of lncRNA CYTOR/miR-193b-3p/HMGB1 axis in regulating the proliferation and apoptosis of T cell acute lymphoblastic leukemia MOLT-4 cell

作　　者：杨春燕[1] 王海燕[1] 黄倩[1] 程盼盼[1] 李玲[1] 陈璐璐[1] 刘海惠 张顥 YANG Chunyan;WANG Haiyan;HUANG Qian;CHENG Panpan;LI Ling;CHEN Lulu;LIU Haihui;ZHANG Hao(Department of Hematology,the Affiliated Hospital of Jining Medical College,Jining,Shandong,272001)

机构地区：[1]济宁医学院附属医院血液科,山东济宁272001

出　　处：《实用临床医药杂志》2022年第21期90-97,102,共9页Journal of Clinical Medicine in Practice

基　　金：山东省济宁市重点研发计划项目(2021YXNS014)。

摘　　要：目的 探讨lncRNA CYTOR对T细胞急性淋巴细胞白血病(T-ALL)细胞增殖和凋亡的影响及可能机制。方法 体外培养人正常骨髓基质细胞系HS-5和T-ALL细胞系(CCRF-CEM、MOLT-4、CEM-C1),采用逆转录-实时定量聚合酶链反应(RT-qPCR)法检测细胞中lncRNA CYTOR、miR-193b-3p和HMGB1 mRNA表达;采用蛋白质印迹(Western blot)法检测细胞中HMGB1蛋白表达。分别转染si-CYTOR、miR-193b-3p mimics、si-HMGB1及共转染si-CYTOR与anti-miR-193b-3p至MOLT-4细胞,以CCK-8法检测细胞增殖,以流式细胞术检测细胞凋亡,以Western blot法检测细胞Bax和Bcl-2、cleaved-caspase3蛋白表达。通过双荧光素酶报告基因实验验证lncRNA CYTOR与miR-193b-3p、miR-193b-3p与HMGB1的调控关系。结果 与HS-5细胞比较,T-ALL细胞系(CCRF-CEM、MOLT-4、CEM-C1)中lncRNA CYTOR、HMGB1 mRNA及其蛋白表达均升高,miR-193b-3p表达降低,差异有统计学意义(P<0.05)。下调lncRNA CYTOR、上调miR-193b-3p或下调HMGB1后,MOLT-4细胞增殖受到抑制,Bcl-2蛋白表达降低,凋亡率和细胞中Bax、cleaved-caspase3蛋白表达升高,差异有统计学意义(P<0.05)。lncRNA CYTOR竞争性吸附miR-193b-3p,HMGB1是miR-193b-3p的靶基因。上调HMGB1表达可逆转lncRNA CYTOR对T-ALL细胞MOLT-4增殖和凋亡的作用。敲低miR-193b-3p可逆转lncRNA CYTOR对MOLT-4细胞增殖和凋亡的作用。结论 lncRNA CYTOR在T-ALL细胞系中表达上调,其可能通过竞争性吸附miR-193b-3p进而上调HMGB1的表达来促进T-ALL细胞增殖,并阻碍细胞凋亡,其有可能成为T-ALL治疗的分子靶点。Objective To investigate the effect of lncRNA CYTOR on the proliferation and apoptosis of T cell acute lymphoblastic leukemia(T-ALL) cells and its possible mechanism. Methods Human normal bone marrow stromal cell lines HS-5 and T-ALL cell lines(CCRF-CEM, MOLT-4 and CEM-C1) were cultured in vitro, and then the expression of lncRNA CYTOR, miR-193 b-3 p and HMGB1 mRNA in the cells were detected by reverse transcription quantitative real-time polymerase chain reaction(RT-qPCR), and the expression of HMGB1 protein in the cells were detected by western blot. MOLT-4 cells were transfected with si-CYTOR, miR-193 b-3 p mimics, si-HMGB1, or co-transfected with si-CYTOR and anti-miR-193 b-3 p, respectively. And then CCK-8 was used to detect cell proliferation. Flow cytometry was used to detect cell apoptosis, and western blot was used to detect the protein expression of Bax, cleaved-caspase3 and Bcl-2 in cells. The dual luciferase reporter gene experiment was used to verify the regulatory relationship between lncRNA CYTOR and miR-193 b-3 p,miR-193b-3p and HMGB1. Results Compared with HS-5 cells,the expressions of lncRNA CYTOR and HMGB1 mRNA and its protein in T-ALL cell lines( CCRF-CEM,MOLT-4 and CEM-C1) were significantly increased,the expression of miR-193b-3p were significantly decreased( P < 0. 05).After down-regulating lncRNA CYTOR,up-regulating miR-193b-3p,or down-regulating HMGB1,MOLT-4 cell proliferation was inhibited,the expression of Bcl-2 protein in cells significantly decreased,and the rate of apoptosis and the protein expression of Bax,cleaved-caspase3 in cells were significantly increased( P < 0. 05). LncRNA CYTOR competitively adsorbed miR-193b-3p,and HMGB1 was the target gene of miR-193b-3p. Up-regulation of HMGB1 expression reversed the effects of lncRNA CYTOR on the proliferation and apoptosis of MOLT-4 in T-ALL cells. Knockdown of miR-193b-3p reversed the effects of lncRNA CYTOR on the proliferation and apoptosis of MOLT-4cell. Conclusion The expression of lncRNA CYTOR is up-regulated in T-ALL cell lines.

关 键 词：T细胞急性淋巴细胞白血病 lncRNA CYTOR miR-193b-3p HMGB1 细胞增殖 凋亡 

分 类 号：R733.7[医药卫生—肿瘤] R446[医药卫生—临床医学]