circ_0080145 Enhances Imatinib Resistance of Chronic Myeloid Leukemia by Regulating miR-326/PPFIA1 Axis
circ_0080145 Enhances Imatinib Resistance of Chronic Myeloid Leukemia by Regulating miR-326/PPFIA1 Axis
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DOI：
10.1089/cbr.2020.3600
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发表时间：
2020-06-27
影响因子：
3.4
通讯作者：
Jia, Xizhen
中科院分区：
文献类型：
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作者：
Che, Hong;Ding, Hong;Jia, Xizhen
背景：获得性多药耐药性常常被认为是包括慢性粒细胞白血病（CML）在内的恶性肿瘤患者化疗失败的原因。在本研究中，作者研究了环状 RNA 0080145 (circ_0080145) 在 CML 伊马替尼 (IM) 耐药中的作用。 材料和方法：应用定量实时聚合酶链反应测量 circ_0080145、microRNA-326 (miR-326) 和 PTPRF 相互作用蛋白 α 1 (PPFIA1) mRNA 的表达。采用3-(4,5-二甲基-2-噻唑基)-2,5-二苯基-2-H-四唑溴化物(MTT)法测定IM和细胞增殖的半数抑制浓度(IC50)。利用流式细胞术分析来评估细胞凋亡。使用特定试剂盒检查葡萄糖摄取和乳酸产生的水平。通过蛋白质印迹测定检测蛋白质水平。通过双荧光素酶报告基因测定验证了 miR-326 与 circ_0080145 或 PPFIA1 之间的靶向关系。构建小鼠异种移植模型来研究circ_0080145在体内的作用。结果：circ_0080145在IM耐药CML患者和细胞中表达上调。 circ_0080145 沉默可抑制 IM 耐药、细胞生长和糖酵解，并在体外诱导 IM 耐药 CML 细胞凋亡。此外，circ_0080145 敲低可阻断体内肿瘤生长和 IM 耐药性。 miR-326 是 circ_0080145 的靶标，抑制 miR-326 可恢复 circ_0080145 沉默对细胞进展和 IM 耐药的影响。此外，PPFIA1是miR-326的靶基因。在 IM 耐药 CML 细胞中，PPFIA1 过表达消除了 miR-326 对 IM 耐药、细胞生长和糖酵解的抑制作用以及对细胞凋亡的促进作用。 结论：circ_0080145 通过调节 miR-326/PPFIA1 轴促进 IM 耐药，这可能为 CML 治疗提供新途径。
Background: Acquired multidrug resistance is often blamed for the failure of chemotherapy in patients with malignant tumors, including chronic myeloid leukemia (CML). In this study, the authors investigated the role of circular RNA 0080145 (circ_0080145) in imatinib (IM) resistance of CML.Materials and Methods: Quantitative real-time polymerase chain reaction was applied to measure the expression of circ_0080145, microRNA-326 (miR-326), and PTPRF interacting protein alpha 1 (PPFIA1) mRNA. 3-(4, 5-dimethyl-2-thiazolyl)-2, 5-diphenyl-2-H-tetrazolium bromide (MTT) assay was used to determine the half maximal inhibitory concentration (IC50) of IM and cell proliferation. Flow cytometry analysis was utilized to assess cell apoptosis. The levels of glucose uptake and lactate production were examined using specific kits. Protein levels were detected through western blot assay. The targeting relationship between miR-326 and circ_0080145 or PPFIA1 was verified by dual-luciferase reporter assay. The murine xenograft model was constructed to investigate the effect of circ_0080145 in vivo.Results: circ_0080145 was upregulated in IM-resistant CML patients and cells. circ_0080145 silencing suppressed IM resistance, cell growth, and glycolysis and induced apoptosis in IM-resistant CML cells in vitro. Moreover, circ_0080145 knockdown blocked tumor growth and IM resistance in vivo. miR-326 was a target of circ_0080145, and miR-326 inhibition restored the effects of circ_0080145 silencing on cell progression and IM resistance. In addition, PPFIA1 was a target gene of miR-326. The suppressive roles in IM resistance, cell growth and glycolysis, and the promotional role in apoptosis mediated by miR-326 were abolished by PPFIA1 overexpression in IM-resistant CML cells.Conclusion: circ_0080145 contributes to IM resistance via modulating miR-326/PPFIA1 axis, which might provide a novel avenue for CML therapy.