Department of Haematooncology and Bone Marrow Transplantation, Medical University of Lublin, Staszica 11, 20-081, Lublin, Poland.
Ann Hematol. 2011 Feb;90(2):173-83. doi: 10.1007/s00277-010-1045-7. Epub 2010 Aug 17.
In this study, we attempted to assess the interactions of resveratrol, a natural compound present in various plant species, with the purine analogues fludarabine and cladribine in terms of their effects on DNA damage and apoptosis in chronic lymphocytic leukemia (CLL) cells. The experiments were performed ex vivo using short-term cell cultures of blood and bone marrow cells from newly diagnosed untreated patients. We analyzed the expression of active caspase-3 and the BCL-2/BAX ratio as markers of apoptosis and the expression of phosphorylated histone H2AX (γH2AX) and activated ATM kinase, which are reporters of DNA damage. The results of our study revealed that resveratrol induced apoptosis in CLL cells in a tumor-specific manner but did not affect non-leukemic cells, and apoptosis was associated with a decreased BCL2/BAX ratio. Here, we report for the first time that both resveratrol + fludarabine and resveratrol + cladribine caused a higher rate of apoptosis in comparison to the rate caused by a single drug. The percentage of apoptotic cells induced by resveratrol alone was higher in the group of patients with better prognostic markers than in those with worse prognostic markers. However, the rates of apoptosis caused by resveratrol combined with purine analogues were independent of ZAP-70 and CD38 expression and the clinical state of the disease; they were only dependent on the presence of high-risk cytogenetic abnormalities. We also observed an increase in γH2AX expression together with a rise in activated ATM in most of the analyzed samples. The obtained results indicate that resveratrol might warrant further study as a new therapeutic option for CLL patients. This naturally occurring substance may be used as a single agent, especially in older persons for whom there are some limitations for the use of aggressive treatment. On the other hand, a lower purine analogue dose could potentially be used in combination with resveratrol because of their combined effect. One of the mechanisms of action of resveratrol is the induction of DNA damage, which ultimately leads to apoptosis.
在这项研究中，我们试图评估白藜芦醇（一种存在于各种植物物种中的天然化合物）与嘌呤类似物氟达拉滨和克拉屈滨之间的相互作用，以评估它们对慢性淋巴细胞白血病（CLL）细胞中 DNA 损伤和凋亡的影响。实验在体外进行，使用新诊断的未经治疗的患者的血液和骨髓细胞的短期细胞培养物进行。我们分析了活性 caspase-3 的表达和 BCL-2/BAX 比值作为凋亡的标志物，以及磷酸化组蛋白 H2AX（γH2AX）和激活的 ATM 激酶的表达，它们是 DNA 损伤的报告者。我们的研究结果表明，白藜芦醇以肿瘤特异性的方式诱导 CLL 细胞凋亡，但不影响非白血病细胞，凋亡与 BCL2/BAX 比值降低有关。在这里，我们首次报道，与单一药物相比，白藜芦醇+氟达拉滨和白藜芦醇+克拉屈滨均导致更高的凋亡率。与预后不良标志物患者相比，白藜芦醇单独诱导的凋亡细胞百分比在预后较好标志物患者中更高。然而，白藜芦醇与嘌呤类似物联合诱导的凋亡率与 ZAP-70 和 CD38 表达和疾病临床状态无关；它们仅取决于存在高风险细胞遗传学异常。我们还观察到大多数分析样本中γH2AX 表达增加，同时激活的 ATM 增加。研究结果表明，白藜芦醇可能需要进一步研究，作为 CLL 患者的新治疗选择。这种天然存在的物质可以单独使用，特别是对于老年人，因为他们的侵袭性治疗有一些限制。另一方面，由于联合作用，可以潜在地降低嘌呤类似物的剂量并与白藜芦醇联合使用。白藜芦醇的作用机制之一是诱导 DNA 损伤，最终导致凋亡。
