Sydney Cord and Marrow Transplant Facility, Centre for Children's Cancer and Blood Disorders, Sydney Children's Hospital, Sydney, Australia.
Exp Hematol. 2010 Oct;38(10):908-921.e1. doi: 10.1016/j.exphem.2010.06.001. Epub 2010 Jun 9.
The objective of this study was to investigate the effect of small molecule inhibitors of glycogen synthase kinase-3β (GSK-3β) on leukemia cell growth and survival.
Analysis of cytotoxicity and cell proliferation was conducted using the MTS assay, cell-cycle analysis, and division tracking. Apoptosis was investigated by Annexin-V/7-aminoactinomycin D and caspase-3 expression. The effect of GSK-3β inhibitors was also tested in vivo in an animal model of leukemia. Gene expression analysis was performed to identify the genes modulated by GSK-3β inhibition in leukemia cells.
GSK-3β inhibitors suppress cell growth and induce apoptosis in seven leukemia cell lines of diverse origin, four acute myeloid leukemia, one myelodysplastic syndrome, and one acute lymphoblastic leukemia samples. GSK-3β inhibitors are cytotoxic for rapidly dividing clonogenic leukemia blasts, and higher doses of the inhibitors are needed to eliminate primitive leukemia progenitor/stem cells. Slow cell-division rate, low drug uptake, and interaction with bone marrow stroma make leukemia cells more resistant to apoptosis induced by GSK-3β inhibitors. Global gene expression analysis combined with functional approaches identified multiple genes and specific signaling pathways modulated by GSK-3β inhibition. An important role for Bcl2 in the regulation of apoptosis induced by GSK-3β inhibitors was defined by expression analysis and confirmed by using pharmacological inhibitors of the protein. In vivo administration of GSK-3β inhibitors delayed tumor formation in a mouse leukemia model. GSK-3β inhibitors did not affect hematopoietic recovery following irradiation.
Our data support further evaluation of GSK-3β inhibitors as promising novel agents for therapeutic intervention in leukemia and warrant clinical investigation in leukemia patients.
本研究旨在探讨小分子糖原合酶激酶-3β（GSK-3β）抑制剂对白血病细胞生长和存活的影响。
采用 MTS 检测法、细胞周期分析和分裂追踪法分析细胞毒性和细胞增殖，用 Annexin-V/7-氨基放线菌素 D 和 caspase-3 表达分析检测细胞凋亡。还在白血病动物模型中测试了 GSK-3β 抑制剂的作用。进行基因表达分析，以确定 GSK-3β 抑制在白血病细胞中调节的基因。
GSK-3β 抑制剂可抑制七种不同起源的白血病细胞系的细胞生长并诱导其凋亡，其中包括四种急性髓性白血病、一种骨髓增生异常综合征和一种急性淋巴细胞性白血病样本。GSK-3β 抑制剂对快速分裂的克隆性白血病母细胞具有细胞毒性，需要更高剂量的抑制剂才能消除原始白血病祖细胞/干细胞。低分裂率、低药物摄取率以及与骨髓基质的相互作用使白血病细胞对 GSK-3β 抑制剂诱导的凋亡具有更强的抵抗力。通过基因表达分析结合功能方法确定了多个受 GSK-3β 抑制调节的基因和特定信号通路。通过表达分析确定了 Bcl2 在 GSK-3β 抑制剂诱导的细胞凋亡调节中的重要作用，并通过使用该蛋白的药理学抑制剂进行了验证。GSK-3β 抑制剂在体内给药可延缓小鼠白血病模型中的肿瘤形成。GSK-3β 抑制剂不影响照射后造血恢复。
我们的数据支持进一步评估 GSK-3β 抑制剂作为治疗白血病的有前途的新型药物，并需要在白血病患者中进行临床研究。
