代谢变化推动了肾细胞癌(RCC)的发展，而黑暗时分分泌的神经荷尔蒙褪黑激素(MLT)对RCC细胞生长及其潜在机制的影响仍不明确。我们通过使用UPLC-MS/MS进行代谢组学分析来检测代谢物浓度，使用Seahorse细胞外流量分析仪来确定氧气消耗率。我们观察到MLT在体外和体内都有效抑制了RCC细胞的生长。此外，MLT增加了ROS水平，抑制了抗氧化酶活性并诱导了细胞凋亡。此外，MLT治疗上调了TCA循环关键代谢物的水平，同时降低了有氧糖酵解产物的水平，导致氧气消耗率、ATP产量和细胞膜电位的增加。此外，MLT治疗还抑制了RCC细胞中Akt、mTOR和p70 S6 Kinase的磷酸化以及HIF-1α/VEGFA的表达；这些效应通过NAC（ROS抑制剂）逆转。相反，MLT与舒尼替尼在RCC细胞中协同抑制细胞生长，并抵消舒尼替尼引起的Warburg效应。总之，我们的结果表明，MLT治疗逆转了Warburg效应并促进细胞内ROS产生，从而抑制了Akt/mTOR/S6K信号通路的活化，诱导细胞凋亡，并与舒尼替尼协同抑制RCC细胞的生长。总体而言，本研究为我们深入了解MLT在RCC细胞中的抗肿瘤作用机制提供了新的见解，并提出MLT可能是治疗RCC的有希望的治疗方法。

Metabolic alteration drives renal cell carcinoma (RCC) development, while the impact of melatonin (MLT), a neurohormone secreted during darkness, on RCC cell growth and underlying mechanisms remains unclear.We detected concentration of metabolites through metabolomic analyses using UPLC-MS/MS, and the oxygen consumption rate was determined using the Seahorse Extracellular Flux analyzer.We observed that MLT effectively inhibited RCC cell growth both in vitro and in vivo. Additionally, MLT increased ROS levels, suppressed antioxidant enzyme activity, and induced apoptosis. Furthermore, MLT treatment upregulated key TCA cycle metabolites while reducing aerobic glycolysis products, leading to higher oxygen consumption rate, ATP production, and membrane potential. Moreover, MLT treatment suppressed phosphorylation of Akt, mTOR, and p70 S6 Kinase as well as the expression of HIF-1α/VEGFA in RCC cells; these effects were reversed by NAC (ROS inhibitors). Conversely, MLT synergistically inhibited cell growth with sunitinib and counteracted the Warburg effect induced by sunitinib in RCC cells.In conclusion, our results indicate that MLT treatment reverses the Warburg effect and promotes intracellular ROS production, which leads to the suppression of Akt/mTOR/S6K signaling pathway, induction of cell apoptosis, and synergistically inhibition of cell growth with sunitinib in RCC cells. Overall, this study provides new insights into the mechanisms underlying anti-tumor effect of MLT in RCC cells, and suggests that MLT might be a promising therapeutic for RCC.