Surgery is the common treatment for early lung cancer with multiple pulmonary nodules, but it is often accompanied by the problem of significant malignancy of other nodules in non-therapeutic areas. In this study, we found that a combined treatment of local radiofrequency ablation (RFA) and melatonin (MLT) greatly improved clinical outcomes for early lung cancer patients with multiple pulmonary nodules by minimizing lung function injury and reducing the probability of malignant transformation or enlargement of nodules in non-ablated areas. Mechanically, as demonstrated in an associated mouse lung tumor model, RFA not only effectively remove treated tumors but also stimulate antitumor immunity, which could inhibit tumor growth in non-ablated areas. MLT enhanced RFA-stimulated NK activity and exerted synergistic antitumor effects with RFA. Transcriptomics and proteomics analyses of residual tumor tissues revealed enhanced oxidative phosphorylation and reduced acidification as well as hypoxia in the tumor microenvironment, which suggests reprogrammed tumor metabolism after combined treatment with RFA and MLT. Analysis of residual tumor further revealed the depressed activity of MAPK, NF-kappa B, Wnt, and Hedgehog pathways and upregulated P53 pathway in tumors, which was in line with the inhibited tumor growth. Combined RFA and MLT treatment also reversed the Warburg effect and decreased tumor malignancy. These findings thus demonstrated that combined treatment of RFA and MLT effectively inhibited the malignancy of non-ablated nodules and provided an innovative non-invasive strategy for treating early lung tumors with multiple pulmonary nodules. Trial registration: www.chictr.org.cn, identifier ChiCTR2100042695, http://www.chictr.org.cn/showproj.aspx?proj=120931.

手术是早期肺癌多发肺结节的常见治疗方法，但常伴有非治疗区其他结节明显恶性的问题。在这项研究中，我们发现局部射频消融 (RFA) 和褪黑激素 (MLT) 的联合治疗通过最大限度地减少肺功能损伤和降低恶性转化或扩大的可能性，极大地改善了多发肺结节的早期肺癌患者的临床结局。未消融区域的结节。在机械上，如相关的小鼠肺肿瘤模型所证明的，RFA 不仅可以有效去除治疗过的肿瘤，还可以刺激抗肿瘤免疫，从而抑制非消融区域的肿瘤生长。MLT 增强 RFA 刺激的 NK 活性并与 RFA 发挥协同抗肿瘤作用。残留肿瘤组织的转录组学和蛋白质组学分析显示，肿瘤微环境中的氧化磷酸化增强，酸化降低以及缺氧，这表明在 RFA 和 MLT 联合治疗后，肿瘤代谢发生了重编程。对残留肿瘤的分析进一步揭示了肿瘤中MAPK、NF-kappa B、Wnt和Hedgehog通路的活性降低和P53通路的上调，这与抑制的肿瘤生长一致。RFA 和 MLT 联合治疗也逆转了 Warburg 效应并降低了肿瘤恶性程度。因此，这些研究结果表明，RFA 和 MLT 的联合治疗有效地抑制了未消融结节的恶性，并为治疗具有多个肺结节的早期肺肿瘤提供了一种创新的非侵入性策略。试用注册：www.chictr.org.cn，