越来越多的证据表明昼夜紊乱对癌症进展具有致癌作用。为了测试通过饮食策略瞄准昼夜节律是否抑制肺癌进展，我们采用 6 小时限时喂养 (TRF) 范式来阐明 TRF 是否以及如何影响肺癌进展。本研究使用了多种肺癌细胞系、两只异种移植小鼠模型和化学处理的小鼠肺癌模型。构建了稳定的 TIM 敲低和 TIM 过表达 A549 细胞。通过集落形成、EdU 增殖、伤口愈合、Transwell 迁移、流式细胞仪和 CCK8 测定来确定体外癌症行为。免疫荧光、病理学检查和靶向代谢组学也用于肿瘤细胞和组织。使用mCherry-GFP-LC3质粒检测自噬流。我们首次发现，与正常随意喂养相比，6小时TRF抑制肺癌进展，并重新编程了参与糖酵解的代谢物或基因的节律和昼夜节律在肿瘤中。 TRF干预后，发现五个肺癌相关时钟基因中只有timeless（TIM）基因能够使肿瘤细胞的节律与肿瘤组织的节律一致。此外，我们证明 TRF 的抗肿瘤作用部分是由 TIM 的节律性下调和随后的自噬激活介导的。 TRF 与 TIM 抑制相结合，进一步增强了抗肿瘤效果，与异种移植模型中化疗的治疗效果相当。六小时 TRF 抑制肺癌进展并重塑昼夜代谢，部分是通过 TIM 的节律性下调和随后的节律介导的。自噬的上调。© 2023。作者。

Accumulating evidence has suggested an oncogenic effect of diurnal disruption on cancer progression. To test whether targeting circadian rhythm by dietary strategy suppressed lung cancer progression, we adopted 6-h time-restricted feeding (TRF) paradigm to elucidate whether and how TRF impacts lung cancer progression.This study used multiple lung cancer cell lines, two xenograft mouse models, and a chemical-treated mouse lung cancer model. Stable TIM-knockdown and TIM-overexpressing A549 cells were constructed. Cancer behaviors in vitro were determined by colony formation, EdU proliferation, wound healing, transwell migration, flow cytometer, and CCK8 assays. Immunofluorescence, pathology examinations, and targeted metabolomics were also used in tumor cells and tissues. mCherry-GFP-LC3 plasmid was used to detect autophagic flux.We found for the first time that compared to normal ad libitum feeding, 6-h TRF inhibited lung cancer progression and reprogrammed the rhythms of metabolites or genes involved in glycolysis and the circadian rhythm in tumors. After TRF intervention, only timeless (TIM) gene among five lung cancer-associated clock genes was found to consistently align rhythm of tumor cells to that of tumor tissues. Further, we demonstrated that the anti-tumor effect upon TRF was partially mediated by the rhythmic downregulation of the TIM and the subsequent activation of autophagy. Combining TRF with TIM inhibition further enhanced the anti-tumor effect, comparable to treatment efficacy of chemotherapy in xenograft model.Six-hour TRF inhibits lung cancer progression and reshapes circadian metabolism, which is partially mediated by the rhythmic downregulation of the TIM and the subsequent upregulation of autophagy.© 2023. The Author(s).