College of Pharmacy, Kyung Hee University, Seoul 130-701, Republic of Korea.
Biol Pharm Bull. 2010;33(6):1054-9. doi: 10.1248/bpb.33.1054.
Although donepezil, a potent acetylcholinesterase (AChE) inhibitor, has been used to treat Alzheimer's disease (AD) due to its neuroprotective effects, its mode of action to inhibit the growth of cancer cells is poorly understood. In the present study, we investigated the pro-apoptotic activities of donepezil in HL-60 human promyelocytic leukemia cells and the underlying molecular mechanism involved. It was found that donepezil induced the apoptosis of HL-60 and U937 cells in a dose- and time-dependent manner, as evidenced by the formation of DNA fragmentation and the accumulation of positive cells for Annexin V. In addition, the activations of caspase-8, -9, and -3 were significantly increased 36 h after donepezil treatment. Furthermore, the broad caspase inhibitor (z-VAD-fmk) blocked donepezil-induced apoptosis. In addition, donepezil was found to cause the loss of mitochondrial membrane potential (DeltaPsi(m)), to increase the release of cytochrome c to the cytosol, and to alter the expressions of Bcl-2 family proteins. Taken together, these results demonstrate for the first time that donepezil displayed an induction of apoptosis in HL-60 cells via a mitochondria-mediated caspase-dependent pathway.
尽管多奈哌齐（一种强效的乙酰胆碱酯酶抑制剂）因其神经保护作用而被用于治疗阿尔茨海默病（AD），但其抑制癌细胞生长的作用机制尚不清楚。在本研究中，我们研究了多奈哌齐对 HL-60 人早幼粒细胞白血病细胞的促凋亡作用及其相关的分子机制。结果表明，多奈哌齐以剂量和时间依赖的方式诱导 HL-60 和 U937 细胞凋亡，这表现在 DNA 片段的形成和 Annexin V 阳性细胞的积累上。此外，多奈哌齐处理 36 小时后，caspase-8、-9 和 -3 的活性显著增加。此外，广谱半胱氨酸蛋白酶抑制剂（z-VAD-fmk）阻断了多奈哌齐诱导的细胞凋亡。此外，多奈哌齐还导致线粒体膜电位（ΔPsi(m)）丧失，细胞色素 c 向细胞质释放增加，并改变 Bcl-2 家族蛋白的表达。总之，这些结果首次表明，多奈哌齐通过线粒体依赖性半胱氨酸蛋白酶依赖途径诱导 HL-60 细胞凋亡。
