Laboratory of Cutaneous Physiopathology, San Gallicano Dermatological Institute, IRCCS, 00144 Rome, Italy.
Tumor Immunology and Immunotherapy Unit, Department of Research Advanced Diagnostic and Technological Innovation, Regina Elena National Cancer Institute, IRCCS, 00144 Rome, Italy.
Cells. 2023 Apr 6;12(7):1099. doi: 10.3390/cells12071099.
Stimulation of melanocytes and murine melanoma cells with αMSH plus the PI3K inhibitor LY294002 resulted in ROS increase, oxidative DNA damage, and pigment retention. We performed cellular and molecular biology assays (Western blot, FACS, immunofluorescence analysis, scratch assay) on murine and human melanoma cells. Treatment with αMSH plus LY294002 altered cortical actin architecture. Given that cytoskeleton integrity requires energy, we next evaluated ATP levels and we observed a drop in ATP after exposure to αMSH plus LY294002. To evaluate if the αMSH-activated PI3K pathway could modulate energy metabolism, we focused on glucose uptake by analyzing the expression of the Glut-1 glucose translocator. Compared with cells treated with αMSH alone, those exposed to combined treatment showed a reduction of Glut-1 on the plasma membrane. This metabolic alteration was associated with changes in mitochondrial mass. A significant decrease of the cell migratory potential was also observed. We demonstrated that the αMSH-dependent PI3K pathway acts as a regulator of energy metabolism via glucose uptake, influencing the actin cytoskeleton, which is involved in melanosome release and cell motility. Hence, these results could constitute the basis for innovative therapeutical strategies.
用 αMSH 和 PI3K 抑制剂 LY294002 刺激黑素细胞和鼠黑色素瘤细胞会导致 ROS 增加、氧化 DNA 损伤和色素保留。我们对鼠和人黑色素瘤细胞进行了细胞和分子生物学检测（Western blot、FACS、免疫荧光分析、划痕实验）。用 αMSH 和 LY294002 处理会改变皮质肌动蛋白的结构。由于细胞骨架的完整性需要能量，我们接下来评估了 ATP 水平，结果发现暴露于 αMSH 和 LY294002 后 ATP 水平下降。为了评估 αMSH 激活的 PI3K 途径是否可以调节能量代谢，我们专注于通过分析葡萄糖转运体 Glut-1 的表达来研究葡萄糖摄取。与仅用 αMSH 处理的细胞相比，联合处理的细胞在质膜上的 Glut-1 减少。这种代谢变化与线粒体质量的变化有关。还观察到细胞迁移能力的显著下降。我们证明，αMSH 依赖性 PI3K 途径通过葡萄糖摄取作为能量代谢的调节剂发挥作用，影响肌动蛋白细胞骨架，这与黑色素体释放和细胞迁移有关。因此，这些结果可能为创新的治疗策略奠定基础。
