College of Laboratory Medicine, Dalian Medical University, Dalian 116044, 9 Lushunnan Road Xiduan, Dalian, 116044, Liaoning Province, China.
Department of Clinical Laboratory, Beijing Hospital of Traditional Chinese Medicine Affiliated to Capital University of Medicine Sciences, Beijing, 100010, China.
J Exp Clin Cancer Res. 2019 May 16;38(1):199. doi: 10.1186/s13046-019-1208-x.
Noncoding RNAs, including microRNAs (miRNAs) and long non-coding RNAs (lncRNAs) are becoming key parts in the development of multidrug resistance (MDR) in T-cell acute lymphoblastic leukemia (T-ALL). Abnormal expression in sialyated N-glycans have been observed in MDR leukemia. However, the role of sialylation regulated MDR remains poorly understood. The aim of this work is to analyze the alternation of N-glycans in T-ALL MDR.
Here, mass spectrometry (MS) is analyzed to screen the N-glycan profiles from ALL cell line CR and adriamycin (ADR)-resistant CR (CR/A) cells. The expression of sialyltransferase (ST) genes in T-ALL cell lines and bone marrow mononuclear cells (BMMCs) of T-ALL patients were analyzed using qRT-PCR. Functionally, T-ALL cell proliferation and MDR are detected through CCK8 assay, colony formation assay, western blot and flow cytometry. RIP assay and Dual-luciferase reporter gene assay confirm the binding association between ZFAS1 and miR-150. Xenograft nude mice models are used to determine the role of ST6GAL1 in vivo.
Elevated expression of α2, 6-sialyltransferase 1 (ST6GAL1) has been detected. The altered level of ST6GAL1 was corresponding to the drug-resistant phenotype of T-ALL cell lines both in vitro and in vivo. ZFAS1/miR-150/ST6GAL1 axis was existed in T-ALL cell lines. MiR-150 was downregulated and inversely correlated to ST6GAL1 expression. ZFAS1 was a direct target of miR-150 and positively modulated ST6GAL1 level by binding miR-150. ZFAS1/miR-150/ST6GAL1 axis functioned to regulate ADR-resistant cell growth and apoptosis. Besides, EGFR was demonstrated to be a substrate of ST6GAL1, and the sialylated EGFR had an impact on the PI3K/Akt pathway.
Results suggested that ZFAS1/miR-150/ST6GAL1 axis involves in the progression of T-ALL/MDR further mediates sialylated EGFR via PI3K/Akt pathway. This work might have an application against T-ALL MDR.
非编码 RNA 包括 microRNAs（miRNAs）和长非编码 RNA（lncRNAs），它们已成为 T 细胞急性淋巴细胞白血病（T-ALL）多药耐药（MDR）发展的关键部分。在耐药性白血病中已经观察到唾液酸化 N-聚糖的异常表达。然而，唾液酸化调节的 MDR 作用仍知之甚少。本研究旨在分析 T-ALL MDR 中 N-聚糖的变化。
采用质谱（MS）分析筛选 ALL 细胞系 CR 和阿霉素（ADR）耐药 CR（CR/A）细胞的 N-聚糖图谱。采用 qRT-PCR 分析 T-ALL 细胞系和 T-ALL 患者骨髓单核细胞（BMMC）中唾液酸转移酶（ST）基因的表达。通过 CCK8 测定、集落形成测定、western blot 和流式细胞术检测 T-ALL 细胞增殖和 MDR。RIP 测定和双荧光素酶报告基因测定证实 ZFAS1 与 miR-150 之间的结合关系。利用裸鼠异种移植模型在体内确定 ST6GAL1 的作用。
检测到α2,6-唾液酸转移酶 1（ST6GAL1）的表达升高。ST6GAL1 的改变水平与体外和体内 T-ALL 细胞系的耐药表型相对应。ZFAS1/miR-150/ST6GAL1 轴存在于 T-ALL 细胞系中。miR-150 下调并与 ST6GAL1 表达呈负相关。ZFAS1 是 miR-150 的直接靶标，通过结合 miR-150 正向调节 ST6GAL1 水平。ZFAS1/miR-150/ST6GAL1 轴通过调节 ADR 耐药细胞的生长和凋亡来发挥作用。此外，EGFR 被证明是 ST6GAL1 的底物，唾液酸化的 EGFR 对 PI3K/Akt 通路有影响。
结果表明，ZFAS1/miR-150/ST6GAL1 轴参与 T-ALL/MDR 的进展，并通过 PI3K/Akt 通路进一步介导唾液酸化的 EGFR。这项工作可能在对抗 T-ALL MDR 方面具有应用价值。
