UNASSIGNED: Quantitative real-time polymerase chain reaction (qRT-PCR) assay was conducted for detecting the expression of circ_0001602, CCND3, microRNA-192-5p (miR-192-5p), and Zinc Finger and BTB Domain-Containing Protein 20 (ZBTB20) mRNA. RNase R assay and Actinomycin D assay were implemented to determine the characteristics of circ_0001602. Cell counting Kit-8 (CCK-8) assay was performed to evaluate cell proliferation. Flow cytometry was employed for assessing cell cycle distribution and apoptosis. Dual-luciferase reporter assay and RIP assay were utilized for confirming the interactions between miR-192-5p and circ_0001602 or ZBTB20.
UNASSIGNED: Circ_0001602 and ZBTB20 were upregulated and miR-192-5p level was reduced in AML tissues and cells. Depletion of circ_0001602 repressed cell proliferation and induced cell cycle arrest and apoptosis in AML cells. Functionally, circ_0001602 was identified to be the sponge of miR-192-5p, and miR-192-5p silence restored the suppressive effects of circ_0001602 knockdown on AML cell progression. Furthermore, ZBTB20 was a target of miR-192-5p, and ZBTB20 overexpression neutralized the miR-192-5p-mediated inhibiting actions on the malignant phenotypes of AML cells. Besides, circ_0001602 could sponge miR-192-5p to positively regulate ZBTB20 expression.
UNASSIGNED: Circ_0001602 contributed to AML cell development at least partially through modulating the miR-192-5p/ZBTB20 axis, which provided new insights for AML treatment.
■进行定量实时聚合酶链反应(qRT-PCR)检测circ_0001602,CCND3,microRNA-192-5p(miR-192-5p)的表达,和锌指和含BTB结构域的蛋白20(ZBTB20)mRNA。实施RNaseR测定和放线菌素D测定以确定circ_0001602的特征。进行细胞计数试剂盒-8(CCK-8)测定以评估细胞增殖。流式细胞术用于评估细胞周期分布和细胞凋亡。双荧光素酶报告基因测定和RIP测定用于确认miR-192-5p与circ_0001602或ZBTB20之间的相互作用。
■Circ_0001602和ZBTB20上调,miR-192-5p水平在AML组织和细胞中降低。circ_0001602的耗尽抑制AML细胞的细胞增殖并诱导细胞周期停滞和凋亡。功能上,circ_0001602被鉴定为miR-192-5p的海绵,miR-192-5p沉默恢复了circ_0001602敲低对AML细胞进展的抑制作用。此外,ZBTB20是miR-192-5p的靶标,和ZBTB20过表达中和miR-192-5p介导的对AML细胞恶性表型的抑制作用。此外,circ_0001602可以海绵化miR-192-5p正向调节ZBTB20的表达。
■Circ_0001602至少部分通过调节miR-192-5p/ZBTB20轴促进AML细胞发育,这为AML治疗提供了新的见解。