Graduate Institute of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
Department of Radiation Oncology, Faculty of Medicine, College of Medicine, Kaohsiung Medical University, Kaohsiung 80708, Taiwan.
Biomolecules. 2020 May 17;10(5):777. doi: 10.3390/biom10050777.
Withaferin A (WFA) has been reported to inhibit cancer cell proliferation based on high cytotoxic concentrations. However, the low cytotoxic effect of WFA in regulating cancer cell migration is rarely investigated. The purpose of this study is to investigate the changes in migration and mechanisms of oral cancer Ca9-22 cells after low concentrations of WFA treatment. WFA under 0.5 μM at 24 h treatment shows no cytotoxicity to oral cancer Ca9-22 cells (~95% viability). Under this condition, WFA triggers reactive oxygen species (ROS) production and inhibits 2D (wound healing) and 3D cell migration (transwell) and Matrigel invasion. Mechanically, WFA inhibits matrix metalloproteinase (MMP)-2 and MMP-9 activities but induces mRNA expression for a group of antioxidant genes, such as nuclear factor, erythroid 2-like 2 (), heme oxygenase 1 (), glutathione-disulfide reductase (), and NAD(P)H quinone dehydrogenase 1 ()) in Ca9-22 cells. Moreover, WFA induces mild phosphorylation of the mitogen-activated protein kinase (MAPK) family, including extracellular signal-regulated kinases 1/2 (ERK1/2), c-Jun N-terminal kinase (JNK), and p38 expression. All WFA-induced changes were suppressed by the presence of ROS scavenger -acetylcysteine (NAC). Therefore, these results suggest that low concentration of WFA retains potent ROS-mediated anti-migration and -invasion abilities for oral cancer cells.
维甲酰胺 A（WFA）已被报道基于高细胞毒性浓度抑制癌细胞增殖。然而，WFA 对调节癌细胞迁移的低细胞毒性作用很少被研究。本研究的目的是研究低浓度 WFA 处理后口腔癌细胞 Ca9-22 迁移的变化和机制。24 小时处理时，浓度低于 0.5 μM 的 WFA 对口腔癌细胞 Ca9-22 无细胞毒性（~95%活力）。在这种情况下，WFA 触发活性氧（ROS）的产生，并抑制 2D（划痕愈合）和 3D 细胞迁移（transwell）和 Matrigel 侵袭。在机制上，WFA 抑制基质金属蛋白酶（MMP）-2 和 MMP-9 的活性，但诱导一组抗氧化基因的 mRNA 表达，如核因子，红系 2 样 2（）、血红素加氧酶 1（）、谷胱甘肽二硫化物还原酶（）和 NAD（P）H 醌氧化还原酶 1（）在 Ca9-22 细胞中。此外，WFA 诱导丝裂原激活蛋白激酶（MAPK）家族成员，包括细胞外信号调节激酶 1/2（ERK1/2）、c-Jun N-末端激酶（JNK）和 p38 的轻度磷酸化。所有 WFA 诱导的变化都被 ROS 清除剂乙酰半胱氨酸（NAC）的存在所抑制。因此，这些结果表明，低浓度的 WFA 保留了对口腔癌细胞具有强大的 ROS 介导的抗迁移和抗侵袭能力。
