Ge Haitao, Kong Xiuqin, Shi Limei, Hou Lijuan, Liu Zhili, Li Ping
Ministry of Education and Department of Pharmacognosy, China Pharmaceutical University, Nanjing, PR China.
Cell Biol Int. 2009 Mar;33(3):402-10. doi: 10.1016/j.cellbi.2009.01.014. Epub 2009 Feb 7.
Various polyunsaturated fatty acids, especially gamma-linolenic acid (GLA), inhibit the growth of a variety of tumor cells. Some evidence indicates that polyunsaturated fatty acid can kill cells by apoptosis. In the current study, we tested the apoptotic effect of GLA on human chronic myelogenous leukemia K562 cells. GLA induced K562 cell death in a dose-dependent manner. Typical apoptotic nuclei were shown by staining of K562 cells with DNA-binding fluorochrome Hoechst 33342, characterized by chromatin condensation and nuclear fragmentation. Flow cytometric analysis also demonstrated that GLA caused dose-dependent apoptosis of K562 cells. The apoptosis could be inhibited by a pancaspase inhibitor (z-VAD-fmk), suggesting the involvement of caspases. Further, release of cytochrome c, activation of caspase-3 and cleavage of PARP were found in GLA-induced apoptosis. GLA treatment could also elevate lipid peroxidation in K562 cells, and antioxidant alpha-tocopherol could reverse the cytotoxicity of GLA. The saturated fatty acid SA, which did not exhibit significant increase in lipid peroxidation, also did not induce cytotoxicity. Intracellular GSH was also determined, and there was no marked change of GSH levels in cells after incubation with GLA compared with the control. These results demonstrate that GLA could induce apoptosis in K562 cells. Apoptosis is mediated by release of cytochrome c, activation of caspase-3. Lipid peroxidation may play a role in GLA cytotoxicity.
多种多不饱和脂肪酸,尤其是γ-亚麻酸(GLA),可抑制多种肿瘤细胞的生长。一些证据表明,多不饱和脂肪酸可通过凋亡杀死细胞。在本研究中,我们测试了GLA对人慢性粒细胞白血病K562细胞的凋亡作用。GLA以剂量依赖性方式诱导K562细胞死亡。用DNA结合荧光染料Hoechst 33342对K562细胞进行染色,显示出典型的凋亡细胞核,其特征为染色质浓缩和核碎裂。流式细胞术分析也表明,GLA导致K562细胞发生剂量依赖性凋亡。泛半胱天冬酶抑制剂(z-VAD-fmk)可抑制这种凋亡,提示半胱天冬酶参与其中。此外,在GLA诱导的凋亡过程中,发现了细胞色素c的释放、半胱天冬酶-3的激活和PARP的裂解。GLA处理还可提高K562细胞中的脂质过氧化水平,抗氧化剂α-生育酚可逆转GLA的细胞毒性。饱和脂肪酸SA未表现出脂质过氧化的显著增加,也未诱导细胞毒性。还测定了细胞内谷胱甘肽(GSH),与对照组相比,用GLA孵育后细胞中GSH水平没有明显变化。这些结果表明,GLA可诱导K562细胞凋亡。凋亡是由细胞色素c的释放、半胱天冬酶-3的激活介导的。脂质过氧化可能在GLA的细胞毒性中起作用。