Department of Cytobiochemistry, Faculty of Biology and Environmental Protection, University of Łódź, 90-236 Łódź, Poland.
Oncol Rep. 2013 May;29(5):2065-71. doi: 10.3892/or.2013.2301. Epub 2013 Feb 22.
There is a current need for novel therapeutic strategies for the treatment of chronic lymphocytic leukemia (CLL), a still incurable hematological cancer involving mainly deregulated apoptosis. The purpose of the present study was to determine ex vivo the effect of the synthetic statin, atorvastatin, a known cholesterol-lowering drug, on peripheral blood mononuclear cells obtained from CLL patients. Using flow cytometry, we investigated the viability and induction of apoptosis in leukemic cells exposed to statin by the Vybrant apoptosis assay kit #4, compared with untreated control cells. We also examined the expression levels of apoptosis-regulatory proteins (Mcl-1, Bcl-2 and Bax), as well as products of the expression/proteolysis of lamin B, poly(ADP-ribose) polymerase‑1 (PARP‑1) and p27Kip1 by western blot analysis. Moreover, the number of sub-G1 cells and DNA fragmentation in atorvastatin-treated leukemic cells were examined by flow cytometry and agarose gel electrophoresis, respectively. The obtained results indicated that CLL cells ex vivo were extremely sensitive to atorvastatin. The cytotoxic effect of this statin was caused by the induction of apoptosis in the leukemic cells. The induction of apoptosis in the drug-treated model cells was confirmed by the reduction or proteolysis of apoptotic markers, such as PARP-1, lamin B and p27Kip1, the increase in the number of sub-G1 cells and DNA ladder formation. During atorvastatin-triggered apoptosis, changes in the expression levels of mitochondrial outer membrane permeability regulatory proteins of the Bcl-2 family were also observed. Ex vivo promising data indicate the strong cytotoxic and pro-apoptotic potential of atorvastatin against leukemic cells, but not normal cells. The obtained data suggest that atorvastatin be considered as a therapeutic option for the treatment of CLL.
目前需要新的治疗策略来治疗慢性淋巴细胞白血病（CLL），这是一种仍无法治愈的血液系统癌症，主要涉及失调的细胞凋亡。本研究的目的是确定合成他汀类药物阿托伐他汀对来自 CLL 患者的外周血单个核细胞的体外作用。通过流式细胞术，我们使用 Vybrant 凋亡检测试剂盒 #4 研究了他汀类药物对白血病细胞活力和凋亡诱导的影响，与未处理的对照细胞进行比较。我们还通过 Western blot 分析检查了凋亡调节蛋白（Mcl-1、Bcl-2 和 Bax）以及核层蛋白 B、多聚（ADP-核糖）聚合酶-1（PARP-1）和 p27Kip1 的表达/蛋白水解产物的表达水平。此外，通过流式细胞术和琼脂糖凝胶电泳分别检查阿托伐他汀处理的白血病细胞中 sub-G1 细胞的数量和 DNA 片段化。结果表明，CLL 细胞在体外对阿托伐他汀极其敏感。这种他汀类药物的细胞毒性作用是通过诱导白血病细胞凋亡引起的。药物处理模型细胞中凋亡的诱导通过凋亡标志物（如 PARP-1、核层蛋白 B 和 p27Kip1 的减少或蛋白水解、sub-G1 细胞数量的增加和 DNA 梯形成）得到证实。在阿托伐他汀触发的凋亡过程中，还观察到 Bcl-2 家族线粒体外膜通透性调节蛋白的表达水平发生变化。体外有前途的数据表明阿托伐他汀对白血病细胞具有很强的细胞毒性和促凋亡作用，但对正常细胞没有作用。这些数据表明，阿托伐他汀可被视为治疗 CLL 的一种治疗选择。
