Department of Biomedical Science and Environmental Biology, PhD Program of Life Science, College of Life Science, Kaohsiung Medical University, Kaohsiung, Taiwan.
School of Dentistry, College of Dental Medicine, Kaohsiung Medical University, Kaohsiung, Taiwan.
Oxid Med Cell Longev. 2021 Mar 2;2021:6667355. doi: 10.1155/2021/6667355. eCollection 2021.
We previously found that marine sponge-derived manoalide induced antiproliferation and apoptosis of oral cancer cells as well as reactive species generations probed by dichloro-dihydrofluorescein diacetate (DCFH-DA) and MitoSOX Red. However, the sources of cellular and mitochondrial redox stresses and the mutual interacting effects between these redox stresses and apoptosis remain unclear. To address this issue, we examined a panel of reactive species and used the inhibitors of cellular reactive species (-acetylcysteine (NAC)), mitochondrial reactive species (MitoTEMPO), and apoptosis (Z-VAD-FMK; ZVAD) to explore their interactions in manoalide-treated oral cancer Ca9-22 and CAL 27 cells. Hydroxyl (˙OH), nitrogen dioxide (NO˙), nitric oxide (˙NO), carbonate radical-anion (CO ), peroxynitrite (ONOO), and superoxide (O ) were increased in oral cancer cells following manoalide treatments in terms of fluorescence staining and flow cytometry. Cellular reactive species (˙OH, NO , ˙NO, CO , and ONOO) as well as cellular and mitochondrial reactive species (O ) were induced in oral cancer cells following manoalide treatment for 6 h. NAC, MitoTEMPO, and ZVAD inhibit manoalide-induced apoptosis in terms of annexin V and pancaspase activity assays. Moreover, NAC inhibits mitochondrial reactive species and MitoTEMPO inhibits cellular reactive species, suggesting that cellular and mitochondrial reactive species can crosstalk to regulate each other. ZVAD shows suppressing effects on the generation of both cellular and mitochondrial reactive species. In conclusion, manoalide induces reciprocally activation between cellular and mitochondrial reactive species and apoptosis in oral cancer cells.
我们之前发现海洋海绵来源的 manoalide 诱导口腔癌细胞的增殖和凋亡，以及二氯二氢荧光素二乙酸酯（DCFH-DA）和 MitoSOX Red 探测到的活性物质生成。然而，细胞和线粒体氧化还原应激的来源以及这些氧化还原应激与凋亡之间的相互作用仍不清楚。为了解决这个问题，我们检查了一组活性物质，并使用细胞活性物质的抑制剂（N-乙酰半胱氨酸（NAC））、线粒体活性物质（MitoTEMPO）和凋亡（Z-VAD-FMK；ZVAD）来探讨它们在 manoalide 处理的口腔癌细胞 Ca9-22 和 CAL 27 中的相互作用。荧光染色和流式细胞术显示 manoalide 处理后口腔癌细胞中羟基（˙OH）、二氧化氮（NO˙）、一氧化氮（˙NO）、碳酸根自由基阴离子（CO ）、过氧亚硝酸盐（ONOO）和超氧阴离子（O ）增加。 manoalide 处理 6 小时后，口腔癌细胞中诱导产生细胞活性物质（˙OH、NO 、˙NO、CO 和 ONOO）以及细胞和线粒体活性物质（O ）。NAC、MitoTEMPO 和 ZVAD 在 annexin V 和 pancaspase 活性测定中抑制 manoalide 诱导的凋亡。此外，NAC 抑制线粒体活性物质，MitoTEMPO 抑制细胞活性物质，表明细胞和线粒体活性物质可以相互作用调节彼此。ZVAD 对细胞和线粒体活性物质的生成均有抑制作用。总之，manoalide 诱导口腔癌细胞中细胞和线粒体活性物质之间的相互激活以及凋亡。
