Fomin A S, Koval' O A, Semenov D V, Potapenko M O, Kuligina E V, Kit Iu Ia, Rikhter V A
Bioorg Khim. 2012 Jan-Feb;38(1):92-8. doi: 10.1134/s1068162012010086.
Earlier we isolated and characterized human milk pro-apoptotic peptide - lactaptin and generated its engineered analog - RL2. It was shown that both lactaptin and RL2 are capable to induce apoptotic death of MCF-7 cells. In this report we have analyzed biochemical markers of RL2 induced MCF-7 apoptosis. The activation of initiator and effector caspases as well as mitochondrial membrane potential and cytoplasm membrane changes were analyzed using flow cytometry and Western-blot methods. We have found that RL2 induced apoptotic death of MCF-7 cells was accompanied by PS exposure on the plasma membrane surface. It also was shown that RL2 has induced dissipation of mitochondrial membrane potential and resulted in activation of initiator caspases 8, 9 and effector caspase 7.
此前我们分离并鉴定了人乳促凋亡肽——乳铁蛋白,并制备了其工程类似物——RL2。结果表明,乳铁蛋白和RL2均能够诱导MCF-7细胞发生凋亡性死亡。在本报告中,我们分析了RL2诱导MCF-7细胞凋亡的生化标志物。使用流式细胞术和蛋白质免疫印迹法分析了起始和效应半胱天冬酶的激活以及线粒体膜电位和细胞质膜的变化。我们发现,RL2诱导的MCF-7细胞凋亡性死亡伴随着质膜表面磷脂酰丝氨酸(PS)的暴露。还发现RL2可诱导线粒体膜电位耗散,并导致起始半胱天冬酶8、9和效应半胱天冬酶7的激活。