Circ_0013745靶向调控miR-126-5p对慢性髓系白血病细胞K562的迁移、侵袭和增殖的影响  

Effects of circ_0013745 targeting miR-126-5p on migration,invasion and proliferation of chronic myeloid leukemia K562 cells

作　　者：焦卫云[1] 刘媛媛[1] 鲍扬漪[1] JIAO Weiyun;LIU Yuanyuan;BAO Yangyi(Department of hematopathology,the First People′s Hospital of Hefei City,Hefei 230000,China)

机构地区：[1]合肥市第一人民医院血液内科,合肥230000

出　　处：《新疆医科大学学报》2023年第1期63-68,73,共7页Journal of Xinjiang Medical University

基　　金：安徽省中央引导地方科技发展专项项目(YN2019032)。

摘　　要：目的探讨circ_0013745靶向调控miR-126-5p对慢性髓系白血病(CML)细胞K562增殖和侵袭的影响。方法收集2019年7月—2021年1月在合肥市第一人民医院诊断治疗的CML患者与健康捐赠者外周血样本30对,利用实时荧光定量PCR(RT-qPCR)检测CML患者中circ_0013745表达量。通过平板克隆实验和细胞计数试剂盒8(Cell counting kit-8,CCK-8)检测circ_0013745对K562细胞增殖能力的影响。Transwell实验检测K562细胞的迁移和侵袭能力。Starbase数据库预测与circ_0013745结合的miRNA,通过双荧光素酶报告系统验证circ_0013745是否与miR-126-5p结合,并分为mimic NC与circ_0013745-WT/circ_0013745-MUT共转染组、miR-126-5p mimic与circ_0013745-WT/circ_0013745-MUT共转染组。通过挽救实验对细胞增殖能力进行检测。结果RT-qPCR检测结果显示,CML患者的外周血中circ_0013745的表达量显著高于健康人表达量(t=7.369,P<0.001)。平板克隆实验结果显示,si-circ_0013745组的K562细胞克隆个数较si-NC组的细胞克隆个数显著降低(P<0.001)。CCK-8增殖实验结果显示,si-circ_0013745组细胞增殖能力较si-NC组细胞增殖能力显著降低(t=8.413,P<0.01)。Transwell迁移实验结果显示,si-circ_0013745组细胞迁移能力低于si-NC组迁移能力(t=7.739,P<0.01)。Transwell侵袭实验结果显示,si-circ_0013745组的侵袭细胞个数较si-NC组显著降低(t=4.679,P<0.01)。Starbase数据库分析结果显示,miR-126与circ_00137453’UTR结合。双荧光素酶报告基因实验显示,与NC组比较,miR-126-5p mimic与circ_0013745-WT共转染组的荧光素酶活性显著降低(t=11.130,P<0.001),而miR-126-5p mimic与circ_0013745-Mut共转染组的荧光素酶活性无明显变化(t=0.316,P>0.05)。敲除circ_0013745后,miR-126-5p的表达量显著上升(P<0.001)。挽救实验结果显示,与si-circ_0013745组细胞增殖率比较,si-circ_0013745+miR-126-5p inhibitor组细胞增殖率增高(t=7.175,P<0.01)。结论Circ_0013745靶向调控miR-126-5p促进CML细胞K562Objective To investigate the effects of circ_0013745 targeting miR-126-5p on proliferation and invasion of chronic myeloid leukemia(CML)K562 cells.Methods 30 pairs of peripheral blood samples of CML patients diagnosed and treated in the hospital from July 2019 to January 2021 and peripheral blood samples of healthy donors were collected.Real-time fluorescence quantitative PCR(RT-qPCR)was used to detect the expression of circ_0013745 in patients with chronic myeloid leukemia.The effects of circ_0013745 on K562 cell proliferation were detected by plate cloning experiment and Cell Counting Kit 8(CCK-8).Transwell assay was used to detect migration and invasion of K562 cells.The miRNA binding to circ_0013745 was predicted by Starbase database,and whether circ_0013745 bound to miR-126-5p was verified by dual lucifase reporting system.And it divided into co-transfection groups with mimic NC and circ_0013745-WT/circ-0013745-MUT,miR-126-5p mimic and co-transfection groups with circ_0013745-WT/circ_0013745-MUT.The proliferation ability of the cells were tested by salvage experiment.Results RT-qPCR showed that the expression of circ_0013745was significantly higher than that of healthy patients in CML patients(t=7.369,P<0.001).The results of plate cloning experiments showed that the number of K562 cell clonesin si-circ_0013745 group was significantly lower than the number of cell clones in si-NC group(P<0.001).The results of CCK-8 proliferation assay showed that the proliferation capacity of si-circ_0013745 group was significantly lower than si-NC group(t=8.413,P<0.01).The results of migration assay showed that the cell migration capacity of si-circ_0013745 group was lower than that of the si-NC group(t=7.739,P<0.01).And results of the invasion assay showed that the number of invasion cells in si-circ_0013745 group was significantly reduced compared with the si-NC group(t=4.679,P<0.01).The results of the starbase database analysis show that,combining miR-126 to circ_00137453’UTR;by the dual-luciferase reporter assay,compar

关 键 词：慢性髓系白血病 细胞侵袭 细胞迁移 细胞增殖 

分 类 号：R733.7[医药卫生—肿瘤]