Li Yi-Ming, Shi Jing, Wu Xu, Luo Zhao-Feng, Wang Feng-Liang, Guo Qing-Xiang
Department of Chemistry, University of Science and Technology of China, Hefei, China.
Cell Biochem Funct. 2009 Oct;27(7):417-23. doi: 10.1002/cbf.1598.
Changes in the free zinc(II) concentration are closely related to cell proliferation and apoptosis, especially during the early apoptotic process. In the present paper, we demonstrated that zinc(II) probe FluoZin-3AM owns sensitive properties to distinguish different stages of apoptotic cell (induced by an anticancer agent, etoposide) according to trace intracellular zinc(II) fluorescence flux. When apoptosis in HeLa or K562 cells was artificially induced, FluoZin-3AM selectively and strongly stained apoptotic cells only at early and middle stages, which was attributed to significantly increased free zinc(II) flux during these stages. This conclusion was further verified by comparing it with the conventional apoptosis detector probe Annexin-V-FITC and PI. Furthermore, FluoZin-3AM was found cell permeable to detect the intracellular zinc(II) fluorescence enhancement to threefolds within 120 s with low cytotoxicity when zinc(II) was incorporated into the cell by zinc(II) ionophore pyrithione. All the above implied that monitoring intracellular zinc fluorescence flux was an effective method to distinguish cell apoptosis from necrosis, and FluoZin-3AM was found to be a suitable probe acting alone to fulfill the work.
游离锌（II）浓度的变化与细胞增殖和凋亡密切相关，尤其是在早期凋亡过程中。在本文中，我们证明了锌（II）探针FluoZin-3AM具有灵敏特性，可根据细胞内微量锌（II）荧光通量来区分凋亡细胞（由抗癌药依托泊苷诱导）的不同阶段。当人工诱导HeLa或K562细胞发生凋亡时，FluoZin-3AM仅在早期和中期选择性地强烈染色凋亡细胞，这归因于这些阶段游离锌（II）通量的显著增加。通过将其与传统凋亡检测探针Annexin-V-FITC和PI进行比较，进一步验证了这一结论。此外，当锌（II）通过锌（II）离子载体吡啶硫酮进入细胞时，发现FluoZin-3AM具有细胞通透性，可在120秒内检测到细胞内锌（II）荧光增强三倍，且细胞毒性较低。上述所有结果表明，监测细胞内锌荧光通量是区分细胞凋亡与坏死的有效方法，并且发现FluoZin-3AM是一种单独完成此项工作的合适探针。
