Department of Pharmacology, Penn State Hershey Cancer Institute, Penn State College of Medicine, 500 University Drive, Hershey, PA 17033, USA.
Chem Biol Interact. 2012 Oct 25;200(1):28-37. doi: 10.1016/j.cbi.2012.08.022. Epub 2012 Sep 13.
Phenylalkyl isoselenocyanate (ISC) compounds were recently designed in our laboratory by incorporating the anticancer element selenium into a panel of phenylalkyl isothiocyanates (ITCs), known to have anticancer properties. A structural activity investigation was carried out to compare the ISC and ITC panels. Cell viability assay and Annexin V staining for apoptosis showed ISC compounds to be more potent in killing A549 lung adenocarcinoma cells. Both ITCs and ISCs were able to deplete reduced glutathione (GSH) in cells, ISCs more rapidly, but ITCs to a greater extent. ISC compounds had a higher rate of reaction to thiol (-SH) groups as determined by pseudo first order kinetics than the corresponding carbon chain length ITC. The equilibrium concentrations of the GSH and protein thiol conjugates did not differ significantly when comparing sulfur to selenium compounds of the same carbon chain length, and did follow the same trend of displaying decreasing reactivity with increasing carbon chain length for both ITCs and ISCs. Furthermore, only ITCs were able to induce cell cycle arrest, suggesting that protein targets inside the cell may differ for the S and Se panels. Finally, the panels were tested for their ability to redox cycle when reacted with GSH to form superoxide and other reactive oxygen species (ROS). ISC compounds showed a much greater ability to redox cycle than corresponding ITCs, and were able to induce higher levels of ROS in A549 cells. Also, the direct pro-apoptotic effects of ISCs and ITCs were inhibited by GSH and potentiated by depletion of intracellular GSH by buthionine sulfoximine. In conclusion, our studies suggest that the redox-cycling capabilities of ISCs and thus generation of higher levels of ROS may be contributing to the increased cytotoxicity of ISC compounds in A549 cells, compared to that of the corresponding ITCs.
苯丙异硒氰酸酯(ISC)化合物是最近由我们实验室设计的,通过将抗癌元素硒引入一组苯丙异硫氰酸酯(ITC)中,这些 ITC 已知具有抗癌特性。进行了一项结构活性研究,以比较 ISC 和 ITC 面板。细胞活力测定和凋亡的 Annexin V 染色表明,ISC 化合物在杀死 A549 肺腺癌细胞方面更有效。ITC 和 ISC 都能够使细胞中的还原型谷胱甘肽(GSH)耗竭,ISC 更快,但 ITC 耗竭程度更大。通过拟一级动力学确定,ISC 化合物与巯基(-SH)基团的反应速率比相应的碳链长 ITC 更高。当比较具有相同碳链长度的硫和硒化合物时,GSH 和蛋白质巯基缀合物的平衡浓度没有显着差异,并且对于 ITC 和 ISC 都遵循随着碳链长度增加而反应性降低的相同趋势。此外,只有 ITC 能够诱导细胞周期停滞,这表明细胞内的蛋白质靶标可能因 S 和 Se 面板而异。最后,当与 GSH 反应形成超氧阴离子和其他活性氧物种(ROS)时,对这些面板进行了测试其氧化还原循环的能力。ISC 化合物比相应的 ITC 具有更大的氧化还原循环能力,并且能够在 A549 细胞中诱导更高水平的 ROS。此外,ISC 和 ITC 的直接促凋亡作用被 GSH 抑制,并通过细胞内 GSH 的耗竭而增强。总之,我们的研究表明,ISC 的氧化还原循环能力以及由此产生的更高水平的 ROS 可能有助于 ISC 化合物在 A549 细胞中的细胞毒性增加,与相应的 ITC 相比。