circ-KEL在急性髓系白血病中的表达及其对白血病细胞的调控作用和机制

Expression of circ-KEL in acute myeloid leukemia and its regulatory mechanisms in leukemic cells

目的探讨circ-KEL在急性髓系白血病(AML)患者中的表达及其对AML细胞的调控作用和机制。方法收集116例AML患者以及40名健康者骨髓标本,分离骨髓单个核细胞,使用实时定量RT-PCR法检测circ-KEL的表达水平并分析其与AML患者临床特征之间的关系。通过生物信息学分析以及双荧光素酶报告基因实验等验证circ-KEL与miR-335-5p/LRG1的靶向关系。运用CCK-8、流式细胞术等方法检测circ-KEL在AML细胞中的生物学作用。结果circ-KEL在AML患者中的表达较正常人群明显增高(-7.117±1.831对-8.669±1.771,P<0.001),circ-KEL高表达患者总生存期(OS)明显短于低表达者(P=0.037)。AML患者接受化疗后circ-KEL表达水平较初诊时下降。同时,circ-KEL可充当miR-335-5p的"海绵",靶向调节LRG1。数据库分析显示miR-335-5p高表达患者预后好,且其与LRG1水平呈负相关。LRG1可促进AML细胞的增殖,抑制其凋亡,在AML患者中表达水平较健康人群也显著升高。circ-KEL可以通过miR-335-5p/LRG1轴发挥对白血病细胞的促增殖以及抑制凋亡作用。结论circ-KEL在AML患者中高表达,与AML患者预后高度相关,通过影响miR-335-5p/LRG1在AML疾病进展中发挥重要作用。

Objective To explore the expression of circ-KEL in patients with acute myeloid leukemia(AML)and the effect and mechanism of circ-KEL on leukemic cells.Methods The expression of circ-KEL was detected by quantitative real-time polymerase chain reaction in bone marrow mononuclear cells collected from 116 patients with AML and 40 healthy donors.The correlation of circ-KEL expression with the clinical characteristics of patients with AML was further systematically analyzed.The modulations among circ-KEL,miR-335-5p,and LRG1 were predicted through bioinformatics analysis and validated by dual luciferase assay.Cell proliferation and apoptosis were detected using CCK8 and flow cytometry.Results The expression of circ-KEL was significantly elevated in patients with AML compared with the healthy controls(Relative expression level,-Δct,AML:-7.117±1.831;control:-8.669±1.771,P<0.001).Moreover,patients with high circ-KEL expression have significantly worse overall survival.The level of circ-KEL in patients with AML was downregulated after chemo-treatment.In addition,circ-KEL could serve as the sponge of miR-335-5p and regulate LRG1.Bioinformatics analysis showed that miR-335-5p correlates with good prognosis and was negatively associated with LRG1.LRG1 could promote cell proliferation and inhibit cell apoptosis.Our results also exhibited the higher expression of LRG1 in patients with AML.Moreover,circ-KEL exerted functional effects via sponging miR-335-5p and regulating LRG1.Conclusion circ-KEL expresses highly in patients with AML and correlates with poor prognosis,suggesting its important role in the genesis and progress of AML.