Department of Pharmaceutical Biology and Biotechnology, Division of Pharmaceutical Biology and Botany, Wroclaw Medical University, 50-556 Wroclaw, Poland.
Department of Molecular and Cellular Biology, Faculty of Pharmacy, Wroclaw Medical University, 50-556 Wroclaw, Poland.
Fitoterapia. 2023 Dec;171:105702. doi: 10.1016/j.fitote.2023.105702. Epub 2023 Oct 15.
Vanicosides A and B isolated from Reynoutria sachalinensis rhizomes are disaccharide phenylpropanoid esters with proven antioxidant activity. Our earlier study showed the cytotoxic activity of vanicosides against melanoma cells, but the mechanism of cell death has not been elucidated. Based on the chemical structure of vanicosides, we proposed that they may induce cell death by generating reactive oxygen species (ROS) into melanoma cells. Moreover, the glucose molecule in their structure can affect the glucose transporters (GLUTs), upregulated in cancer cells. The A375 (melanotic) and C32 (amelanotic) melanoma cell lines were applied. Cell viability assay and ROS-Glo™ assay were performed before and after blocking of Glucose Transporter Type 1 (GLUT1) by WZB117. Fibroblasts and the SKOV-3 line were included in the study to test selectivity in the action of vanicosides and help to elucidate the mechanism of action. Upon incubation with vanicosides, high production of ROS occured, especially inside C32 cells, which was significantly reduced after GLUT-1 blocking. The A375 cells produced less ROS. Melanoma cells were simillary sensitive to the cytotoxic effects of vanicosides, which was clearly enhanced when vanicosides were used together with the WZB117 (GLUT1 inhibitor). The SKOV-3 line and the fibroblasts showed much less sensitivity to the cytotoxicity of vanicosides, also used together with WZB117. Moreover, no significant ROS formation was observed in these lines. The study proved that vanicosides generate ROS inside melanoma cells. These findings suggest that the combination of pro-oxidative acting vanicosides and GLUT1 inhibitors exerts a synergistic cytotoxic effect on melanoma cells.
从薇甘菊根茎中分离得到的香草酸 A 和 B 是二糖苯丙素酯，具有已证实的抗氧化活性。我们之前的研究表明香草酸对黑色素瘤细胞具有细胞毒性作用，但细胞死亡的机制尚未阐明。基于香草酸的化学结构，我们提出它们可能通过向黑色素瘤细胞中生成活性氧（ROS）来诱导细胞死亡。此外，其结构中的葡萄糖分子可以影响癌细胞中上调的葡萄糖转运蛋白（GLUTs）。应用了 A375（色素性）和 C32（非色素性）黑色素瘤细胞系。在使用 WZB117 阻断葡萄糖转运蛋白 1（GLUT1）前后进行了细胞活力测定和 ROS-Glo™测定。将成纤维细胞和 SKOV-3 系纳入研究中，以测试香草酸作用的选择性并帮助阐明作用机制。与香草酸孵育后，ROS 的产生量很高，尤其是在 C32 细胞内，而 GLUT1 阻断后则显著降低。A375 细胞产生的 ROS 较少。黑色素瘤细胞对香草酸的细胞毒性作用同样敏感，当香草酸与 WZB117（GLUT1 抑制剂）一起使用时，这种敏感性明显增强。SKOV-3 系和成纤维细胞对香草酸的细胞毒性也表现出较低的敏感性，同时也与 WZB117 一起使用。此外，在这些细胞系中未观察到明显的 ROS 形成。该研究证明香草酸在黑色素瘤细胞内产生 ROS。这些发现表明，具有促氧化作用的香草酸与 GLUT1 抑制剂的组合对黑色素瘤细胞产生协同的细胞毒性作用。
