School of Pharmacy, Health Science Center, Xi'an Jiaotong University, Xi'an 710061, P.R. China.
State Key Laboratory of Shaanxi for Natural Medicines Research and Engineering, Xi'an 710061, P.R. China.
Curr Cancer Drug Targets. 2022;22(5):404-413. doi: 10.2174/1568009622666220304202116.
T cell acute lymphoblastic leukemia (T-ALL) is an invasive hematological malignant disorder of T cell progenitors. The Janus kinase-signal transducer and activator of transcription (JAK-STAT) signaling pathway plays an important role in the development of T-ALL and in the inhibition of the key molecule, JAK2, and could suppress T-ALL cell proliferation.
The objective of this study was to investigate the in vitro anti-tumor effects of a novel nilotinib derivative, ND-17, on cancer cell lines via its interactions with JAK2.
The effects of ND-17 on cell proliferation and on cell cycle and apoptosis were evaluated using the tetrazolium assay and flow cytometry, respectively. In addition, the ND-17/JAK2 binding interactions were evaluated using surface plasmon resonance and western blot analyses.
ND-17 exerted the greatest inhibitory effects on T-ALL cells amongst all hematological cancer cell lines tested. Flow cytometric analysis indicated that ND-17 blocked the cell cycle at the S phase in T-ALL cells. Nilotinib did not significantly inhibit T-ALL cell growth or regulate the cell cycle. Preliminary investigations revealed that the regulation of cyclin-dependent kinases/cyclins was attributed to ND-17-induced cell cycle arrest. Furthermore, ND-17 could bind to JAK2 with strong affinity, and more importantly, ND-17 bound to the ATP pocket of JAK2 in a manner similar to the potent inhibitor. Thus, ND-17 treatment exhibited a prominent effect in inhibiting the phosphorylation of JAK2 in T-ALL cells. An increase in the phosphorylation of JAK2 was observed in interleukin-6- stimulated Jurkat cells, which was reversed by ND-17 treatment. Meanwhile, the combination of TG- 101348 and ND-17 led to further improvement in inhibiting the phosphorylation of JAK2. Moreover, the transfection and knockdown of JAK2 altered the inhibitory effect of ND-17 on Jurkat cell viability. In addition, ND-17 treatment suppressed the JAK/STAT, phosphatidylinositol-3-kinase/protein kinase B/mechanistic target of rapamycin, and mitogen-activated protein kinase/extracellular signal-regulated protein kinases 1 and 2 signaling pathways.
These findings suggest that ND-17 could be a promising JAK2 inhibitor for the treatment of T-ALL.
T 细胞急性淋巴细胞白血病（T-ALL）是一种侵袭性血液恶性疾病，源于 T 细胞前体细胞。Janus 激酶-信号转导子和转录激活子（JAK-STAT）信号通路在 T-ALL 的发生发展以及关键分子 JAK2 的抑制中发挥着重要作用，可抑制 T-ALL 细胞增殖。
本研究旨在通过与 JAK2 的相互作用，探讨新型尼洛替尼衍生物 ND-17 对癌症细胞系的体外抗肿瘤作用。
采用四唑盐比色法和流式细胞术分别评估 ND-17 对细胞增殖、细胞周期和细胞凋亡的影响。此外，还通过表面等离子体共振和 Western blot 分析评估 ND-17/JAK2 结合相互作用。
ND-17 对所有测试的血液系统恶性肿瘤细胞系中 T-ALL 细胞的抑制作用最大。流式细胞术分析表明，ND-17 可将 T-ALL 细胞的细胞周期阻滞在 S 期。尼洛替尼对 T-ALL 细胞的生长无明显抑制作用，也不能调节细胞周期。初步研究表明，细胞周期蛋白依赖性激酶/细胞周期蛋白的调节归因于 ND-17 诱导的细胞周期阻滞。此外，ND-17 与 JAK2 具有很强的亲和力结合，更重要的是，ND-17 以类似于强效抑制剂的方式与 JAK2 的 ATP 结合口袋结合。因此，ND-17 治疗可显著抑制 T-ALL 细胞中 JAK2 的磷酸化。白细胞介素 6 刺激的 Jurkat 细胞中 JAK2 的磷酸化增加，而 ND-17 治疗可逆转这一现象。同时，TG-101348 和 ND-17 的联合应用可进一步改善 JAK2 的磷酸化抑制作用。此外，JAK2 的转染和敲低改变了 ND-17 对 Jurkat 细胞活力的抑制作用。此外，ND-17 治疗可抑制 JAK/STAT、磷脂酰肌醇 3-激酶/蛋白激酶 B/雷帕霉素靶蛋白、丝裂原激活蛋白激酶/细胞外信号调节激酶 1 和 2 信号通路。
这些发现表明，ND-17 可能是治疗 T-ALL 的有前途的 JAK2 抑制剂。
